1.Establishing a 29 Y-STR Loci Multiplex PCR System.
Xin-jie WANG ; Li-jing LUO ; Lei HUANG ; Xin XU
Journal of Forensic Medicine 2015;31(6):456-461
OBJECTIVE:
To establish a 29 Y-STR loci multiplex PCR system for investigating the genetic polymorphisms and to assess its application value in forensic science.
METHODS:
A multiplex PCR system was established using a five color fluorescence labeling 29 Y-STR loci (DYS456, DYS389 I , DYS437, DYS447, DYS389 11, DYS438, DYS522, DYS460, DYS458, DYS622, DYS390, DYS392, DYS448, DYS449, DYS391, Y-GA TA-H4, DYS388, DYS19, DYS385a/b, DYS527a/b, DYS393, DYS459a/b, DYS635, DYS439, DYS570 and DYS627) for multiple amplification and capillary electrophoresis. And its applicability was validated with genetic polymorphism data of 29 Y-STR of unrelated 2,000 male samples in Shandong Han population.
RESULTS:
A total of 1,981 different haplotypes of 2,000 individuals showed genotype diver- sity between 0.370 0 and 0.965 4. The system provided stable and accurate typing with high sensitivity of 0.05 ng. It satisfied the needs of variety of routine biological samples.
CONCLUSION
The 29 Y-STR loci multiplex PCR system could be applied for actual cases and establishment of Y-STR database. In addition, it has great significance in forensic science practices and related research.
Asian People/genetics*
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China
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Chromosomes, Human, Y
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DNA/isolation & purification*
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Ethnicity/genetics*
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Forensic Genetics/methods*
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Forensic Sciences
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Genetics, Population/methods*
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Haplotypes
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Humans
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Male
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Microsatellite Repeats
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Multiplex Polymerase Chain Reaction/methods*
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Polymorphism, Genetic
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Reproducibility of Results
2.Expression of IL-29 cDNA in cos-7 cells and its anti-HBV activity in vitro
Bing CHEN ; Xin LUO ; Yunxia LU ; Congzhen XU ; Shengquan ZHANG
Chinese Journal of Immunology 1985;0(05):-
Objective: To clone cDNA of human interleukin-29(IL-29) and express it in cos-7 cells, and to study its anti-HBV activity in vitro. Methods: Total RNA was extracted from PBMCs which had been infected with vesicular stomatitis virus(VSV) in vitro.IL-29 cDNA was amplified using one-step RT-PCR technique. The recombinant expressing plasimd of psectagB/His-myc-IL29 was constructed by inserting IL-29 cDNA into the vector and was then transfected into cos-7 cells. Stable expression stains were screened by Hygromycin B and limited dilution method. The target protein was purified through Ni2+-chelating Sepharose Fast Flow. Anti-viral bioactivity of the recombinant IL-29 fusion protein was analyzed through an in vitro model of production of HBV by the HepG2.2.2.15 cell line.ELISA was used for detection of the viral titers in the cell cultural supernants. Results: IL-29 was cloned and stably expressed in cos-7 cells successfully. SDS-PAGE and Western blot analysis showed multiple bands of the target protein with the molecular weights between 20 000 and 33 000, and the major band was located at about 33 000, indicating the fused IL-29 modified by additional glycosylation. The rhIL-29 was shown to dose-dependently inhibit secretion of HBsAg and HBeAg accompanied by the reduction of HBV genomic DNA in the cells tested. The inhibition ratio of HBsAg and HBeAg production was attained 85% at a concentration of 160 ?g/L of rhIL-29. Conclusion: The rhIL-29 with anti-HBV activity has been obtained.
3.Research on the clinical teaching method related to the medical students of obstetrics and Gynecology
Jianping XU ; Yuan SHEN ; Xin LUO ; Qing LI
Chinese Journal of Medical Education Research 2005;0(06):-
Objective To investigate the combination of centralized and decentralized approachesin clinical teaching,in order to improve the teachingway and enhance the clinical quality for the trainees of obstetrics and gynecology.Method Conducting the questionnaire survey with 205 students of obstetrics and gynecology who are interns involving in this clinical method in our hospital. Results Trained by this way,82.9 percent of students believe that they fully understand the true meaning of the clinical work. 86.8 percent and 87.8 percent of students think that the quality of clinical practice is improved and their work ability has been strengthened. 88.3 percent of students agree that good communication between them and patients and their family members is increased. Referring to improvement on the understanding between trainees and their teachers,the number is 85.5 percent .92.6 percent of students think that their analytical sense benefits from this method and 86.8 percent of students agree that this model is useful for their problemsolving skills. Conclusion The combination of centralized and decentralized approach in clinical teaching helps students adapt to the work environment in hospital as soon as possible. Students also understand the truth of the clinical work. This method makes interns and patients as well as family members communicate more easily and the ability to integrate theory with practice is greatly improved.
4.Effect of modified Wuhuang oil on fibroblast growth and proliferation
Guohua XIN ; Xu LUO ; Youlai ZHANG ; Yuanlin ZENG ; Ziyang WAN
Chinese Journal of Tissue Engineering Research 2009;13(50):9988-9991
BACKGROUND: Wuhuang oil has a bacteriostatic action to treat burn wound and promote traumatic healing, but the action on inhibition of scars formation remains poorly understood.OBJECTIVE: To investigate the effects of modified Wuhuang oil at different concentrations and administration times on the growth and proliferation of human fibroblasts in vitro.DESIGN, TIME AND SETTING: Comparison observation regarding cytology in vitro was performed at the Burns Institute in the First Affiliated Hospital of Nanchang University between April 2006 and January 2007.MATERIALS: Prepuce specimens were harvested from patients who underwent circumcision in Department of urinary surgery, at First Affiliated Hospital of Nanchang University and Jiangxi Provincial Children Hospital. All patients aged 2-12 years old, and informed consents were obtained from their relatives. Wuhuang oil and modified Wuhuang oil (water-solubility) were offered by Department of Pharmaceutical Preparation in the First Affiliated Hospital of Nanchang University, China. METHODS: Human fibroblasts cultured in vitro were divided into 2 groups at random, experiment and control. Experiment group was treated with 300 g/L Wuhuang oil, while control group with 300 g/L modified Wuhuang oil. Serum-free culture fluid was used to prepare 6 concentrations of oil solution: 0 (blank control), 100, 150, 200, 250, 300 g/L.MAIN OUTCOME MEASURES: MTT assay was used to determine the growth and proliferation of human fibroblasts at 2, 3, 4, 5,6 days; inhibition rate of cell growth was observed at 2, 4, 6, 8, 10 days.RESULTS: Modified Wuhuang oil (0-300 g/L) concentration positively correlated with inhibition of human fibroblast proliferation;the inhibition was not related to culture time. Modified Wuhuang oil (300 g/L) had the greatest inhibition rate of human fibroblasts at 8-10 days, there were significant differences between experiment group and control group (P < 0.01).CONCLUSION: Modified Wuhuang oil has an effective inhibition on the proliferation of human fibroblasts in vitro, and shows a dose-dependent tendency. Compared with Wuhuang oil, 300 g/L modified Wuhuang oil is superior to suppress the growth of human fibroblasts.
5.Dynamic changes of enzymes activities and growth factors contents in human colostrum
Yide QIN ; Xin LUO ; Denglian HUANG ; Congzhen XU
Chinese Journal of Obstetrics and Gynecology 2001;0(07):-
Objective To study the dynamic changes of the activities of superoxide dismutase (SOD), amylase (AM) and alkaline phosphatase (ALP) and of the contents of epithelium growth factors (EGF) and insulin growth factors (IGF-1) in human colostrum (lactation 1-7 days postpartum). Methods A total of 118 samples of human colostrum were taken from 20 healthy women postpartum. The activities of SOD, AM and ALP in human colostrum were determined by methods of pyrogallol autoxidation, colorimetry of iodine-starch and pyrocatechol phosphate respectively. EGF and IGF-1 contents in human colostrum were measured by radioimmunoassay. Results The SOD activity of human colostrum increased slightly during lactation 1-4 days postpartum, from (18.7?2.2) kU/L to (22.5?2.9) kU/L, then it tended to decrease, being (11.2?2.1) kU/L on the 7th day. The SOD activities in lactation 1-5 days postpartum were significantly higher than that on day 7 ( P
6.Establishing a 29 Y-STR Loci Multiplex PCR System
Xinjie WANG ; Lijing LUO ; Lei HUANG ; Xin XU
Journal of Forensic Medicine 2015;(6):456-461
Objective To establish a 29 Y-STRloci multiplex PC Rsystemfor investigating the genetic polymorphisms and to assess its application value in forensic science. Methods Amultiplex PC Rsystemw as established using a five color fluorescence labeling 29 Y-STRloci (DYS456, DYS389Ⅰ, DYS437, DYS447, DYS389Ⅱ, DYS438, DYS522, DYS460, DYS458, DYS622, DYS390, DYS392, DYS448, DYS449, DYS391, Y-GATA-H4, DYS388, DYS19, DYS385a/b, DYS527a/b, DYS393, DYS459a/b, DYS635, DYS439, DYS570 and DYS627) for multiple amplification and capillary electrophoresis. And its applicability w as validated w ith genetic polymorphismdata of 29 Y-STRof unrelated 2 000 male samples in Shandong Han population. Results Atotal of 1981different haplotypes of 2 000 individuals show ed genotype diver-sity betw een 0.370 0 and 0.965 4. The systemprovided stable and accurate typing w ith high sensitivity of 0.05 ng. It satisfied the needs of variety of routine biological samples. Conclusion The 29 Y-STRloci multiplex PC Rsystemcould be applied for actual cases and establishment of Y-STRdatabase. In addi-tion, it has great significance in forensic science practices and related research.
7.Human IL-21 gene was expressed in Hela cell and it costimulate T cell proliferation in vitro
Shengquan ZHANG ; Bing CHEN ; Xin LUO ; Congzhen XU ; Yunxia LU ;
Chinese Journal of Immunology 1985;0(01):-
Objective:To construct Interleukin 21 (IL 21) expression plasimd pCDNA3.1 IL21 and express it in Hela cell and analysis its acitivity of costimilating T cell proliferation with anti CD3 monoclonal antibody in vitro.Methods:The CDs of IL 21 was amplified by PCR using the pMD IL21 as templet.The expression plasimd pCDNA3.1 IL21 was constructed by inserting the sequence of coding region of the IL 21 into pCDNA3.1/Hismyc B containing CMV promoter and transfected into Hela cells.The stability expression stain was screened by the condition media containing 400 mg/L G418 and cloned through the limited dilution method.The target protein was purified through Ni 2+ chelating Sepharose Fast Flow.The bioactivity was confirmed by MTT method on costimulating the T cell proliferation with anti CD3.SDS PAGE and Western blot analysis the rhIL 21 expressed.Results:hIL 21 was expressed in Hela cell successfully.SDS PAGE analysis showed the IL 21 fusion protein with Mr 18 000 or so was expressed in supernatant of the cells.The rhIL 21 has significant proliferation effect on mature human T cells in the presence of anti CD3 monoantibody.Conclusion:The rhIL 21 with bioactivity has been obtained,which may help researcher study its new function and effects. [
8.The expression of platelet activity in hypertensive patients complicated with ischemic cardiovascular and cerebrovascular disease
Xu LIANG ; Lin-Jie LUO ; Xin-Gen WANG ; Al ET ;
Chinese Journal of Practical Internal Medicine 2001;0(04):-
Objective To observe the change of platelet activity marker in hypertensive patients complicated with ischemic cardiovascular and cerebrovascular disease ,and assess it s value in the happening of thrombotic disease. Method The platelet activity marker (CD62p,CD63) were measured by flow cytometry in 40 hypertensive patients complicated with ischemic cardiovascular and cerebrovascular disease (group C ),46 hypertensive patients not complicated with ischemic cardiovascular and cerebrovascular disease (group B),and 30 cases of healthy control (group A). Comparison between three groups were made. The serum fibrinogen were measured at the same time. Result serum level of CD62p, and CD63 in group B,C are higher than group A.(P
9.Effect of Earlier Rehabilitation on Activities of Daily Living of Patients with Spinal Cord Injury
Zhi-an LUO ; Xu-guang HE ; Jian-xin LI
Chinese Journal of Rehabilitation Theory and Practice 2006;12(3):246-247
ObjectiveTo explore the effect of earlier rehabilitation on activities of daily living(ADL) of patients with spinal cord injury(SCI).Methods50 SCI patients received earlier rehabilitation and improvement of ADL of patients was evaluated.ResultsAfter two months treatment,the scores of Barthel index,functional independence measure(FIM) grade of patients increased significantly compared with that of before treatment(P<0.05) and ADL improved.ConclusionEarlier rehabilitation can improve ADL of SCI patients.
10.Angioimmunoblastic T cell lymphoma with Reed-Sternberg-like cells.
Yan-hui LIU ; Heng-guo ZHUANG ; Dong-lan LUO ; Xin-lan LUO ; Jie XU
Chinese Journal of Pathology 2006;35(4):249-250
Aged
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Antigens, CD20
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analysis
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Gene Rearrangement
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Humans
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Immunoblastic Lymphadenopathy
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genetics
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metabolism
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pathology
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Immunoglobulin Heavy Chains
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genetics
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Immunohistochemistry
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Lymph Nodes
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metabolism
;
pathology
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Lymphoma, T-Cell
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genetics
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metabolism
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pathology
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Male
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Polymerase Chain Reaction
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RNA, Viral
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analysis
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Receptors, Antigen, T-Cell, gamma-delta
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genetics
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Reed-Sternberg Cells
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metabolism
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pathology