Humans ; Siblings ; Forensic Medicine ; Forensic Genetics

Objective To establish a new method for rapid detection of β-thalassemia by investigating six clinical common mutation types.Methods Fifty cases of clinical wild-type samples and 42 cases ofβ-thalassemia samples were collected, and β-globin gene was amplified by PCR.Uniform ligation probe ( ULP) specific probes were designed for hybridization reaction to increase the reaction specificity and real-time PCR was performed to increase the sensitivity.After that, PCR products were verified by agarose electrophoresis.After examining the specificity and sensitivity, Kappa test between LDR-ULP method and reverse dot blot( RDB) method was conducted.Results Hybridization efficiency was improved 2.53 times by LDR-ULP hybridization.Each mutant type showed a significant amplification curve, whereas the wild-type had no significant curve within 40 cycles.The limit of determination of this method was 5 pg.The results of 92 cases of peripheral blood samples detected by the method of LDR-ULP and RDB were completely consistent.Conclusion In this study, a simple, inexpensive, rapid new method to detect β-thalassemia were established.

Objective To establish a homothermal and fast detecting method on pathogenic bacteria by combining recombinase-aid amplification (RAA) with molecular beacon.Methods The establishment of the methodology.Staphylococcus aureus specific primers were designed from the relative region of the staphylococcal protein A (SPA).Asymmetry amplification was optimized by adjusting the primer concentration ratios.The results of amplification and hybridization were visualized and analyzed by agarose gel electrophoresis and fluorescence detection.The sensitivity was identified by detecting dilute positive plasmids.And the specificity was determined using RAA method by detecting 72 pathogenic bacteria,including Staphylococcus aureus and other Staphylococcus spp.from the Department of Clinical Laboratory of Daping Hospital in December 2016.Besides,the Kappa analysis and the clinical diagnosis efficiency were investigated by analyzing 39 extra strains in the laboratory in December 2016.Results When the concentration ratio of restrictive and non-restrictive primer was 1:20,the yield efficiency of single-stranded DNA (ssDNA) reached the peak.And as for the hybridization efficiency,the asymmetry amplification was higher than symmetry amplification.Twenty copies/μl was proposed as the limits of detection by testing dilute plasmids.And the RAA hybridization method could distinguish Staphylococcus aureus with other Staphylococcus spp.Comparing with traditional detection methods with a Kappa index of 0.860,this method shows a good consistency.By analyzing the 111 bacteria,the sensitivity of the method is 92.5％ (37/40),the specificity is 97.2％ (69/71),the positive predictive value is 94.9％ (37/39),the negative predictive value is 95.8％ (69/72),the positive likelihood radio is 33.04,the negative likelihood radio is 0.077,the Youden index is 0.897 and the Kappa index is 0.902.Conclusion Through the combination of asymmetry recombinase-aid amplification optimization and molecular beacon probe,a new method of detecting bacteria DNA with RAA hybridization technique is established,providing the foundation for its clinical application.

Objective: To investigate the awareness of pubertal knowledge and its influencing factors among primary school students in Chinese rural areas, and to provide evidences for improving awareness level and popularizing scientific, reasonable, and suitable sex education. Methods: Sixteen primary schools consisting of rural registered students in Guizhou and Anhui Province were selected by multi-stage cluster sampling method, and a questionnaire survey for 2 506 senior students (grade five and six) was conducted to explore the awareness of pubertal knowledge and its influencing factors among primary school students in Chinese rural areas. Results: The total awareness rate of pubertal knowledge was 62.7% among primary school students in Guizhou and Anhui rural areas, and the awareness rates were relatively low for the questions about the difference between male and female pubertal development and understanding of sexual attitude; female students, only-child students and grade six students got a bit better awareness of pubertal knowledge than male students, non-only-child students and grade five students respectively; grade[OR=0.72(0.61-0.86)], peer acceptance [OR=0.70(0.53-0.96)], family intimacy [OR=0.75(0.63-0.88)] had effect on awareness of pubertal knowledge. Conclusion The awareness of pubertal knowledge among primary school students in Guizhou and Anhui rural areas need further improvement, especially for male students, non-only-child students and grade five students in the questions about the difference between male and female pubertal development and understanding of sexual attitude. It is necessary to improve the awareness of pubertal knowledge by enhancing peer acceptance, promoting family intimacy, and popularizing scientific, reasonable, and suitable sex education.

AIMTo ascertain the bioactivity and to analyse quantificationally the denervating action of botulinum toxin A (BTXA) in gel.

METHODS36 Sprague-Dawley rats were randomized into four groups. In group A - D, the gastrocnemius muscle of one leg was randomly selected to receive injection of BTXA solution 5U in 0.1 ml, BTXA gel 12.5U in 0.1 ml, BTXA gel 5U in 0.1 ml and BTXA gel 2U in 0.1 ml respectively, while the gastrocnemius muscle of other leg was injected with 0.1 ml of saline solution in group A and 0.1 ml of gel in group B to group D as control. Compound muscle action potential (CMAP) of both gastrocnemius muscles were measured and the amplitudes were recorded before injections, and 5 days, 2 weeks, 3 weeks, 1 month, 2 months and 3 months after the injections respectively.

RESULTSThe reduction of CMAP amplitude was significantly different at various time (P < 0.01), and CMAP amplitude decreased significantly after the treatment of BTXA (P < 0.01). The reduction of CMAP amplitude was significantly dif ferent in group A to I) (P < 0.01), and more reduction was found in group A and B (P < 0.01), and the reduction was higher in group C than in group D (P < 0.05). However, there were no significant differences in the reduction of CMAP amplitude between group A and group B.

CONCLUSIONBioactivity of BTXA in gel was showed and the denervating action of BTXA in gel was demonstrated in a dosage and time dependent manner.

Animals ; Botulinum Toxins, Type A ; administration & dosage ; Dosage Forms ; Female ; Gels ; Injections, Intramuscular ; Mice ; Muscle Denervation ; methods ; Muscle, Skeletal ; innervation ; Rats ; Rats, Sprague-Dawley ; Solutions

OBJECTIVETo provide new germplasm materials for breeding new varieties of Pseudostellaria heterophylla.

METHODThe method of single plant selection was adopted, with the comparative experiments being carried out under the same conditions in Shibing county. The 8 plants of Shibing SB-4 were compared respectively with factor analysis for 27 phenotypic traits and 8 yield traits, and single factor variance analysis for the contents of polysaccharides.

RESULTUsing factor analysis, 27 phenotypic traits were classified into 7 principal divisors and 8 yield traits were simplified into 3 principal divisors. The 4 strains of P. heterophylla, ZT-01, ZT-02, ZT-06 and ZT-07, performed better than others in the phenotypic traits, and ZT-01, ZT-02, ZT-03 and ZT-07 in the yield traits. The contents of polysaccharides of ZT-01, ZT-02, ZT-05 and ZT-08 showed significantly higher value.

CONCLUSIONThere is significant difference among the 8 strains of P. heterophylla in phenotypic traits, yield traits and quality traits, making it possible to select certain strains for different purposes. ZT-01 and ZT-02 can be breaded further. ZT-06 and ZT-07 were used as ornamental cultivars for its great phenotypic traits. ZT-03 with good resistance and high yield was taken as resistant variety, and ZT-05 would face next selection on the basis of its high content of polysaccharide.

Breeding ; Caryophyllaceae ; chemistry ; growth & development ; China ; Phenotype ; Polysaccharides ; analysis

Objective: To understand the incidence of school violence and its related factors among senior students in rural primary schools in Guizhou and Anhui provinces in 2016. Methods: From September to November 2016, one county was randomly selected from 47 impoverished counties of Guizhou and one county was randomly selected from 18 impoverished counties of Anhui Province by using multi-stage cluster sampling method. A total of 4 schools were randomly selected from primary schools which had more than 70% students with rural resident registration in each selected county, and 3 classes were randomly selected from fifth and sixth grades of each selected school. All students in each selected class were included in this study. A total of 2 521 students were included, and 2 506 valid questionnaires were collected with an effective response rate about 99.4%. Self-designed questionnaires were used to collect data on individual information, family information, and the incidence of school violence with related factors. Multivariable logistic regression was conducted to analyze related factors of school violence. Results: The incidence of violence in rural primary schools was 50.4% (1 264/2 506) in 2016. The number of students who had physical assault, fought or been bullied was 682 (27.2%), 689 (27.5%) and 764 (30.5%), respectively. The results of multivariable logistic regression showed that the OR (95%CI) values of incidence of school violence for male students was 2.13(1.78-2.54) compared with female students; and the OR(95%CI) values of incidence of school violence for students with high peer acceptance, high family intimacy, more physical activity and mother who had education background of junior high school, senior high school, technical secondary school and junior college, bachelor′s degree or above were 0.39 (0.26-0.57), 0.47 (0.40-0.57), 0.64 (0.52-0.78), 0.79 (0.66-0.96), and 0.65 (0.46-0.92) compared with those with low peer acceptance, low family intimacy, less physical activity and mother who had primary education and below. Conclusion The incidence of violence in rural primary schools is not optimistic. Gender, peer acceptance, family intimacy, physical activity and mother education are related to school violence.

Objective To observe the effect of high-frequency repetitive transcranial magnetic stimulation(rTMS)on hand function in patients after stroke. Methods From June,2016 to September,2017,30 stroke patients were randomly divided into control group(n=30)and experimental group(n=30).Both groups received routine training after sham rTMS or 3 Hz rTMS.Fugl-Meyer Assessment(FMA),modified Ashworth Scale(MAS)and modified Barthel Index(MBI)were used to evaluate the hand function and the activities of daily living before and after treatment. Results Finally 27 patients completed the experiment.After treatment, the FMA score increased in both groups (Z>2.070, P<0.05), and the difference value was higher in the experimental group than in the control group (Z=-2.296,P<0.05);the MAS score improved in the experimental group(Z=-2.456,P<0.05),no difference was found in the control group(Z=-0.816,P>0.05),and the difference value was higher in the experimental group than in the control group(Z=-2.792,P<0.01);the MBI score improved in both groups(t>3.085,P<0.01),howev-er,no difference was found in the MBI score and the difference value between two groups(t<0.246,P>0.05). Conclusion High-frequency rTMS could promote the hand function in patients after stroke.

OBJECTIVETo construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.

METHODSPeripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).

CONCLUSIONThe constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; immunology ; Antibody Specificity ; Arthritis, Rheumatoid ; immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin kappa-Chains ; biosynthesis ; genetics ; Lymphocytes ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection

OBJECTIVETo construct and characterize EGFP reporter gene labeled Sindbis virus (SINV).

METHODSThe reporter gene EGFP was inserted into the genome of infectious clone pBR-XJ160 by using multi-fusion long fragment PCR method. Then apply reverse genetic manipulation technique to rescue and obtain EGFP labeled SINV.

RESULTSWe successively obtained labeled SINV, which has good fluorescent expression characteristics and genetic stability.

CONCLUSIONThe labeled virus can be seen in living cells and living body, and this serves as a good tool for cell and tissue tropism and biological function study of viruses. This study laid a foundation for further studying the cell tropism, biological functions and infection mechanism of SINV.

Base Sequence ; Genes, Reporter ; Green Fluorescent Proteins ; genetics ; Molecular Sequence Data ; Sindbis Virus ; genetics