Preliminary exploration on operation process for autologous ozonized blood transfusion / 中国输血杂志

Autologous ozonized blood transfusion(AOBT) is a therapy of re-transfusion of 100-200 mL of autologous blood after shaking and agitation with appropriate amount of oxygen-ozone in vitro. The oxidation of blood through the strong oxidation of ozone can enhance the non-specific immune response of the body, regulate the internal environment and promote health. This therapy has been increasingly applied in clinical practice, while no unified standard for the operation process in terms of ozone concentration, treatment frequency and treatment course had been established. This operation process of AOBT is primarily explored in order to standardize the operation process and ensure its safety and efficacy.

Association between birth defects and special risk exposures during peri-conception period / 西安交通大学学报(医学版)

Objective To analyze the effect of special risk exposures during periconception period on birth defects of newborns.Methods From Jul.to Dec.2013,the multi-stage stratified random sampling method was adopted.Women of childbearing age between 15 and 49 who were pregnant during 2010 to 2013 in Shaanxi Province were selected as study subjects for investigation on special risk factors exposed during periconception period.The Logistic regression model was adopted to analyze the association between newborns' birth defects and special risk exposures.Results The study included 30 010 women of childbearing age and 29 550 newborns with 572 (193.57/ 10 000) cases of birth defects.After adjusting for demographic factors,the risk factors for birth defects were drinking [OR=2.29,95％ CI (1.22,4.29)] and passive smoking [OR=1.25,95％ CI (1.02,1.53)] during periconception.There was a higher risk of birth defects when exposure to medicine [OR =1.64,95％ CI (1.04,2.61)],pesticides [OR =2.41,95％ CI (1.09,5.35)],biological risk factors [OR-1.64,95％ CI (1.05,2.56)],physical risk factors [OR=1.15,95％ CI (1.13,2.34)] and chemical risk factors [OR =2.36,95％ CI (1.36,4.11)] 3 months both before and after pregnancy.Similarly,after adjusting for demographic factors and behaviors,we found that birth defects were related to antibiotics,salicylates,and antitussive,which could increase the risk of birth defects (P＜0.05).Conclusion Exposure to passive smoking and drinking during periconception and exposure to medicines and pesticides,as well as biological,physical and chemical risk factors 3 months before and after pregnancy could increase the risk of birth defects in newborns.

Association between congenital heart disease and folic acid supplementation during periconceptional period among women of childbearing age in Shaanxi / 西安交通大学学报(医学版)

Objective To explore the association between folic acid supplementation during periconcerptional period and congenital heart disease in newborns to provide scientific evidence for making intervening measures.Methods Using stratified random cluster sampling,a total of 30 counties were sampled from Shaanxi Province.A questionnaire survey was conducted among childbearing-aged women pregnant between January 2010 and November 2013.All of the included women had definite pregnancy outcomes and had signed the consent form.Logistic regression was performed to investigate the association between folic acid supplementation during pregnancy and congenital heart disease in newborns.Results In total,28 354 questionnaires were available for analysis.The overall prevalence of congenital heart disease among live-birth neonates in the present study was 7.3‰.The percentage of childbearing-age women who had taken folic acid supplementation during pregnancy was 64.4％,while only 17.2％ of them took folic acid according to the specification.Taking folic acid regularly during pregnancy was associated with a lower risk of congenital heart disease among the newborns (OR 0.502,95％ CI:0.279 0.902).The multiple-factor analysis results also showed that taking folic acid regularly during periconcerptional period could reduce the risk of congenital heart disease (adjusted OR=0.512,P=0.046) when we controlled the family background factors,mother factors and exposure risk factors during pregnancy.However,no association was found between irregularly taking folic acid during periconcerptional period and the risk of congenital heart disease.Conclusion Taking folic acid according to the specification during periconcerptional period (taking folic acid during 3 months before pregnancy to 3 months after pregnancy with a daily dose of 0.4mg for more than 90 days) may prevent congenital heart disease of newborns.

Two Different Procedures of Blood Donation in Volunteer Donors: A Comparative Study / 中华医院感染学杂志

OBJECTIVE To compare two different procedures of blood donation in volunteer donors,which lead to different discard rates of blood,different donation reaction rates and the satisfaction of the donor agency,so as to seek the better procedure bringing less discard of blood and more convenience for the military donor agency and blood center. METHODS In group A,3 667 donors blood was collected before the tests and retests for transfusion transmitted diseases(TTD) were done.While in group B,4 185 donors were taken blood samples for pre-donation test.The blood collection was performed 4 hours later. RESULTS In group A,3 652 units of blood were collected,of which 69 units were discarded on account of positive results in test and retest.Meanwhile,in group B 3 718 units of blood were collected from the donors who passed the pre-donation test for TTD.As a result,34 units of blood were discarded because of the positive results in retest.The discard rates of blood were 1.89% and 0.91% while the donation reaction rates were 2.22% and 3.98%,respectively.in two procedures.The discard rates of blood in group A were higher than those in group B.But the donation reaction rate in group B was higher than that in group A. CONCLUSIONS The discard rate of blood in the procedure collecting before test is higher than that in the procedure testing before collection.But the donation reaction rate is low and the waiting period for donation is short in the former procedure,which is suitable for low TTD infections population of military agencies.

Association of killer immunoglobulin-like receptors with leukemia / 第三军医大学学报

Objective To study the correlation between the genes of killer immunoglobulin (Ig)-like receptors (KIR) in natural killer cells and leukemia. MethodsPrimers of 16 KIR genes, including KIR2DL1-5, KIR3DL1-3, KIR2DS1-5, KIR3DS1 and 2 pseudogenes 2DP1 and 3DP1, were designed and synthesized. A polymerase chain reaction with sequence-specific primers (PCR-SSP) was developed to detect the mRNA expressions of the 16 genes. Totally 46 healthy individuals and 46 leukemic patients (15 cases of acute myelocytic leukemia, 13 cases of chronic myelocytic leukemia and 18 cases of acute lymphocytic leukemia) were detected and the frequencies of the KIR genes were analyzed. ResultsThe frequency of 2DL4, 3DL2, 3DL3 and 2DP1 in healthy and leukemic patients were 100%, and were detected in all patients and healthy individuals. The other KIR genes had similar frequencies except those of 2DL2 and 2DS2, which were 47.8% of the healthy individuals and 89.1% in leukemic patients. There was a significant difference between the 2 groups.ConclusionThere may be an association between pathogenesis of leukemia and 2DL2 and 2DS2 genes.

Expression of chimeric antigen of HCV with multiple immunodominant epitopes in E .coli and its application for serological tests / 中华传染病杂志

Objective ToexpresschimericantigenofHCVwithmultipleimmunodominantepitopes inE .coliforimprovingthequalityofreagentsforHCVscreening .Methods Thegenefragmentencoding HCVchimericantigenwasobtainedbymolecularcloningmethodandclonedintopQE 30 plasmidforex pressioninE .coliM 15 .TheexpressedchimericantigenwaspurifiedbyNi NTAandcoatedonELISA platestoanalyzeitssensitivityandspecificity .Results Thechimericantigenof 5 30 0 0washighlyex pressed .ELISAassayofserumsamples (including 18HCVpositiveand 17negativesera)indicatedthatthe HCVchimericantigenhadhighsensitivityandspecificity .Conclusions ChimericantigenofHCVwith typicalimmunodominantepitopescanbeusedtodevelopgoodreagentsforHCVimmunoassay .

Cloning and sequencing of partial gene from SENV isolate in China / 军事医学科学院院刊

Objective:To investigate the presence of SENV infection among patients in China,and analyze partial nucleotide sequence of SENV isolated from a patient with non A-G hepatitis.Methods:A nested polymerase chain reaction (PCR) assay with primers from ORF1 of SENV genome was established to detect SENV DNA.The PCR product was cloned and sequenced.Results:SENV DNA was positive in 2 of 7 patients with non A-G hepatitis and TTV negative.Partial gene of a SENV isolate was compared with the corresponding region of SENV isolate(AX025730)from Italy and was found that the nucleotide homology was 90%.Conclusions:The results of this study confirmed the presence of SENV infection in China.The development of a PCR assay for SENV DNA detection and the cloning,sequencing of the SENV isolate have important implication for the diagnosis and epidemiological investigation on SENV infection.

Optimization of method for extraction of template of bacteria for randomly amplified polymorphic DNA / 军事医学科学院院刊

Objective:To obtain optimum template for randomly amplified polymorphic DNA(RAPD). Methods:Four different methods(rod winding, precipitation, boiling and lysis)for extraction of template DNA were used.The length and purity of template DNA and its RAPD profile were observed separately by agarose gel electrophoresis, spectrophotometric scan assays and RAPD reaction.Results:The template DNA (>23 kb) with high purity and the same RAPD profile with 2-4 kb DNA fragments were obtained by both rod winding and precipitation method. However,the template DNA (4 kb and 2 kb,respectively) with break and dispersion and low purity was extracted by method of boiling and lysis, and 600-2 000 bp DNA fragments were seen in the similar RAPD profile.Conclusions: Template DNA extracted by rod winding or precipitation method was optimized for RAPD.

Yeast surface display technology and its applications / 军事医学科学院院刊

Yeast surface display is a new technology developed in recent years,which displays protein of interest on the surface of yeast cell.It can be used to display complex eukaryotic proteins requiring post-translational processing including glycosylation and efficient folding for functional activity. The basis of this technology, its development,methods of screening,its applications and perspective are mainly described in this paper.

Comparison of single factor design and response surface design to optimize the reaction system for randomly amplified polymorphic DNA / 中华检验医学杂志

Objective To obtain optimum scheme on reaction system for randomly amplified polymorphic DNA(RAPD) of Pseudomonas aeruginosa. Methods The optimum reaction systems of RAPD were obtained by single factor design and response surface design(RSD). Results The multiple optimum reaction systems caused by effect of interaction on multifactor were observed in single factor design, and the concentrations of key components of the optimum reaction system with good stability obtained by response surface design were 2.0 mmol/L of Mg 2+ , 0.5 ?mol/L of primer and 0.5 ng/?l of template. Conclusion To optimize the reaction system of RAPD, RSD is simpler, more scientific, and more reasonable than single factor design.

Serial Expression of Human ?,? globin Genes in Escherichia coli / 中国输血杂志

Objective To make an attempt to express serialy the human ?,? globin in Escherichia Coli in order to carry out the research on blood substitute based on the genetic recombination of hemoglobin.Methods The routine molecular biology technology was used.Results The serial genetic fragment of ?,? globins containing the hemoglobin was amplified by PCR.The genetic sequencing showed that there was no unexpected mutation.And then,the serial genes were cloned into the pBV220 expression carrier.Undergoing thermal inducement,the expression product could reach about 20% of total bacterial protein.The expression product assumed the form of inclusion body,and it was vertified by Western Blotting.Conclusion This much can lay the solid foundations for further launching the research on expression and pruification of recombinant human hemoglobin genes.

Cloning and Screening of Mutant Human Hemoglobin Gene(?_(99) ,?_(82) Lys→Cys) / 中国输血杂志

Objective:To mutate the codons which code the lysine at loci ? 99 ,? 82 in human hemoglobin gene to the codons which code the cystein.Methods:The objective gene to be mutated was cloned into pAlter Ex2 plasmis and then was mutated with the site directed mutagenesis mediated by oligonucleotide.Results:The sequencing of clones obtained by preliminary screening showed that the anticipated results were reached without any unexpected mutation.Conclusion:The results lay a foundation for the further development of research on expression and purification of recombinated mutant human hemoglobin gene.

Construction of pIRES-I-A~d?? and stable expression of BALB/c mouse I-A~d?? chain genes in NIH3T3 cell line / 中国免疫学杂志

Objective:To construct pIRES-I-Ad?? bicistronic eukaryotic expression vector. Methods: Total RNA was acquired by TRIZOL method, the genes of I-Ad ?? chains were amplified through RT-PCR, respectively. The target genes were connected to pGEM-T vector and sequenced. Then the target genes were subcloned into pIRES bicistronic eukaiyotic expression vector and NIH3T3 cell line was transfected. Transfectant was screened by G418 antibiotics. Total RNA of transfectant was obtained by TRIZOL method, mRNA of foreign gene was examined by RT-PCR. Flow cytometry( FCM) was used to detennine foreign gene expression in protein level and surface expression in NIH3T3 cell line. Results: Bicistronic eukaryotic expression vector pIRES-I-AdaB was established. Foreign gene in mRNA level in transfectants was examined. Verified Ⅰ-Ad ?? chain wa3 expressed in high level expressed on transfected NIH3T3 cell line surface by FCM. Conclusion:Bicistronic eukaryotic expression vector pIRES-I-Ad ?? was constructed successfully. It is useful for studying antigen presentation and interaction between epitopes and MHC- Ⅱ molecule of BALB/c mouse.

SOLUBLE EXPRESSION OF ? AND ? GENES OF HUMAN HEMOGLOBIN IN ESCHERICHIA COLI / 解放军医学杂志

Fragments of ? and ? genes were cloned into expression plasmid pET-21b. Bacteria harbouring expression plasmid was induced with IPTG,and a band of expressed protein of 16 kD was found in PAGE. They expressed as soluble products. ? expression product reached about 5% of total bacterial protein,? expression product reached about 15% of total bacterial protein. The products were confirmed by Western-blotting.