1.Replacing human peripheral blood leukocytes with LRS-separated leukocytes for scientific research
Yanzheng GU ; Longhai TANG ; Ziyi HUANG ; Weijie LI ; Guangbo ZHANG ; Mingyuan WANG
Chinese Journal of Microbiology and Immunology 2016;36(8):607-614
Objective To investigate the feasibility of using leukocytes that were filtered out by LeukoReduction System ( LRS) to replace conventional human peripheral blood leukocytes in experimental researches and to comparatively analyze the differences between them in vitro biological functions and pheno-types of T cells. Methods Mononuclear cells were isolated from LRS-separated leukocytes and whole blood sample that collected from the same person by using Ficoll. Fluorescence-activated cell sorting ( FACS) was performed to analyze the phenotypes of T cells. CD3+T cells were sorted out by using magnetic beads. The T cells that were collected by using two different ways were incubated with anti-CD3 and anti-CD28 antibodies and IL-2 in vitro for 10 days. Several assays including cell counting, FACS and cytometric beads array ( CBA) were performed to comparatively analyze the differences in biological functions and phenotypes of T cells that were isolated by different methods. Results The phenotypes of T cells isolated from LRS filter and whole blood sample were highly similar at the initial stage. The sorting rate of CD3+T cells form LRS filter reached a high level and met the requirements for experimental researches. No statistically significant differ-ences in cell count, phenotype, expression of costimulatory molecules and cytokine secretion were observed between T cells isolated from LRS filter and whole blood sample. Conclusion This study suggested that the T cells isolated from LRS filter could be used as an alternative to whole blood T cells for fundamental resear-ches since they were similar in cell vitality, phenotype and biological functions. It provided a new way to solve the problem of blood shortage in clinic and scientific research.
2.The analysis of voluntary blood donors with HIV infection in Suzhou city from 2002 to 2013
Li DONG ; Yi CAO ; Hui ZHANG ; Longhai TANG ; Nina JIANG ; Mingyuan WANG ; Yiming JIN
China Modern Doctor 2014;(35):106-108
Objective To analyze the epidemic characteristics of volunteer blood donors with HIV infection in Suzhou city, in order to make countermeasures to recruit appropriate volunteer blood donors. Methods Infectious situation and population characteristics of volunteer blood donors with HIV infection were explored by a retrospective analysis method in Suzhou city between 2012 and 2013. Results Among 755 643 volunteer blood donors from 2002 to 2013, 51 donors were HIV infection, the positive rate was 6.75/100000, and the highest detection rate was found in 2013 (23.25/10万), following as 2012 for 9.71/100000, and the lowest was 1.66/100000. There was a statistically significant differ-ence among these years(χ2=37.96, P<0.01). 42 cases of 51 cases with HIV infection were men, the other 9 were women, sex ratio between male and female was nearly 5:1. The donors with 18 to 30 years old (60.79%) were predominant pop-ulation, unmarried people were as high as 62.75%, the staff were 56.86%, the donors with high school, technical sec-ondary school degrees and following degrees primarily accounted for 50.98%, and individual donors accounted for 70.59% of HIV infections. Conclusion There is an upward trend of HIV infection rate from voluntary blood donors in Suzhou in recent years. Blood collection organization should strengthen the health consultation of voluntary blood donors before the blood donation, especially among young adults, strict screening for HIV, which guarantee for clinical safety of blood transfusion.
3.Anatomical observation on oral part of the facial artery and facial vein and its clinical application
Dongqin YANG ; Lei YU ; Huan BIAN ; Feng TANG ; Yang TAN ; Xueqin BAI ; Yating FU ; Yuexuan HU ; Lan HUANG ; Jing CHEN ; Longhai WU ; Jingping ZHANG ; Yan ZENG ; Xiaobo WANG ; Maocheng RAN
Journal of Regional Anatomy and Operative Surgery 2015;(3):267-269
Objective To observe the oral part of the facial artery and facial vein and to provide anatomical data for clinical applica-tion. Methods The origin, branches, course, diameter, position of oral part of facial artery and facial vein were observed on 32 fixed cada-ves (64 sides). Results The position relation between the facial artery and facial vein is non-constant. Measure the distance from inferior border of mandible to corner of the mouth, angulus mandibulae, mental protuberance midpoint. It is (5. 49 ± 0. 63) cm, (2. 50 ± 0. 89) cm and (6. 20 ± 1. 68) cm in the left side respectively, and (5. 69 ± 0. 72) cm, (2. 56 ± 1. 08) cm and (6. 85 ± 1. 86) cm in the right side re-spectively. The diameter of facial artery in inferior border of mandible is (0. 33 ± 0. 08) cm in the left side and (0. 38 ± 0. 07) cm in the right side;while the diameter of facial vein is (0. 40 ± 0. 12) cm in the left side and (0. 42 ± 0. 18) cm in the right side. The facial artery and facial vein are not concomitant and they are not asymmetry also. The position of superior labial artery arteries is constant, but the position of inferior labial artery arteries have more variations. Conclusion The branches, course, diameter and position of oral part of facial artery and facial vein have a number of variations. The superior labial artery arteries could be positioned more easily than inferior labial artery arter-ies. Being familiar with their distribution is of great importance for clinical application.
4.Investigation on the prevalence of human parvovirus B19 infection among voluntary blood donors
Rong LU ; Xueqiong ZHENG ; Zihao XU ; Longhai TANG ; Mingyuan WANG ; Yiming JIN
Chinese Journal of Blood Transfusion 2023;36(7):622-626
Objective To investigate the infection of human parvovirus B19 in Suzhou voluntary blood donors under the current blood screening model. Methods A total of 893 blood donor samples from September to December 2022 were randomly collected. Samples were tested to determine the seroprevalence (anti-B19 IgG and IgM) of B19 antibodies by enzyme-linked immunosorbent assay(ELISA), and B19 DNA of positive samples was further detected by real-time polymerase chain reaction(PCR) assay. Results Among 893 samples, the total seroprevalence of B19 antibody was 20.7% (185/893), with anti-B19 IgG and IgM positive rate at 19.4% (173/893) and 1.9% (17/893), respectively, showing significant difference (P<0.05). No difference in the positive rates of B19 IgG and IgM (20.1%, 1.5% vs 18.0%,2.6%) was noticed by gender(P>0.05). The prevalence of anti-B19 IgG statistically increased with age (P<0.05), while there was no difference in the prevalence of anti-B19 IgM (P>0.05). No statistical difference was not found in anti-B19 IgG and IgM samples among different blood groups. The anti-B19 IgG in repeated blood donors was higher than that in first-time donors(21.5% vs 15.9%)(P<0.05) while there was no difference in the positive rate of IgM antibodies (P>0.05). Three cases were found to be positive for B19 DNA in the B19 antibody positive samples, with the positive rate at 1.6%(3/185). Conclusion Although the prevalence of B19 infection in Suzhou was lower than that in other areas and was mostly past infection, there was still a certain proportion of persistent infection and acute infection, which posed the potential risk of blood transfusion transmission. Therefore, attention should be paid to blood transfusions, especially for the high-risk and susceptible groups.
5.Causes of HLA-A2 reactive antibodies and analysis of their eplets
Jingjing HUANG ; Longhai TANG ; Yuanling ZUO ; Yan CAO ; Weijie LI ; Min JIANG ; Mingyuan WANG
Chinese Journal of Blood Transfusion 2023;36(6):467-470
【Objective】 To analyze differences of eplets between the patient who generated HLA allele-specific antibodies after platelet transfusion with donors. 【Methods】 The HLA genotypes of the patient and donors were detected by PCR-SBT, and the Luminex single antigen beads coating was used to screen HLA-Ⅰ antibodies in the patient’s serum. HLA Matchmaker was utilized to analyze different amino acids and eplets. 【Results】 The patient carried HLA-A*02∶03 allele, and HLA-A2 antibodies were found in his serum after platelet transfusion (A*02∶01, A*02∶06, and A*02∶07). Sequence alignment showed that the patient′s A*02∶03 has a difference in position 149, which resulted in a different eplet between A*02∶03 and A*02∶01, A*02∶06, A*02∶07 and then induced the production of antibodies. 【Conclusion】 HLA antibodies are specific for HLA epitopes that have structural differences due to amino acid differences between HLA alleles, suggesting that high-resolution typing of HLA-A, -B need to be conducted in patients and donors, and the acceptable mismatch of HLA should be determined based on epitopes rather than antigens, so as to reduce alloimmune response and improve platelet count after transfusion.
6.Platelet antibody of blood donors in Suzhou
Li DONG ; Ming FANG ; Yujue WANG ; Honghong HE ; Hongmei WANG ; Yezhou CHEN ; Feiran WU ; Shengbao DUAN ; Longhai TANG
Chinese Journal of Blood Transfusion 2022;35(8):795-799
【Objective】 To study the incidence and specificity of platelet antibody in blood donors in Suzhou, analyze the distribution characteristics of platelet antibody in blood donors in this area, and explore the significance of platelet antibody detection in blood donors to reduce the adverse reactions toplatelet transfusion in clinical. 【Methods】 Platelet antibody detection was performed in 2178 blood donors in this area by solid-phase immunosorbent assay. The antibody specificity of the positive samples was analyzed by commercial kit, and the anti-CD36 antibody positive samples were further identified by flow cytometry and gene sequencing. 【Results】 Twelve positive samples were detected by platelet antibody screening, with a positive rate of 0.55%(12/2 178), including 5 males (0.33%, 5/2 178)and 7 females(1.06%, 7/2 178). Among the positive samples, anti-HLA-Ⅰ antibody was identified in 2 cases, anti-CD36 antibody in 1 case, and the antibody specificity was not identified in the other 9 cases. In one case, the positive rate of anti-HLA-Ⅰ antibody PRA was 31.31%(31/ 99), which was mainly specific to anti-B15, anti-B35 and anti-B40. The positive rate of anti-HLA-Ⅰ antibody PRA in the other case was 45.45%(45/ 99), which was mainly specific to anti-A2, anti-A11, anti-A24, anti-A29, anti-A33, anti-A66, anti-B15 and anti-B35. The blood donor with anti-CD36 antibody was type I CD36 deficiency, and 329_330delAC mutation occurred in exon 5. 【Conclusion】 Through antibody screening and specificity identification, the positive rate of platelet antibody in females was significantly higher than that in males(P<0.05). In addition to the common anti-HLA-I antibodies, anti-CD36 antibody was also detected in type I CD36 deficient blood donor. Therefore, the detection of platelet antibodies in blood donors is of certain clinical significance to reduce the adverse reactions to blood transfusion caused by antibodies in platelet products.
7.Phenotype screening and gene mutation analysis of platelet CD36 antigen: blood donors in Suzhou area
Honghong HE ; Jing CHEN ; Yezhou CHEN ; Yujue WANG ; Longhai TANG ; Hongmei WANG ; Shaohua DING ; Shuangshi WEI ; Jingjing TIAN ; Shengbao DUAN
Chinese Journal of Blood Transfusion 2021;34(6):585-589
【Objective】 To investigate the expression of CD36 antigen in Suzhou area, analyze the type of antigen deficiency and gene mutation, so as to provide references for the establishment of CD36 negative donor registry in Suzhou. 【Methods】 Anticoagulant whole blood samples (805 cases) were randomly collected from healthy blood donors in Suzhou Blood Center. The expression of CD36 antigen on platelet and monocyte was analyzed by flow cytometry to determine the type of CD36 deficiency. The gene mutation type of platelet CD36 antigen-deficient was performed by genomic DNA sequencing. 【Results】 The CD36 deficiency frequency on platelet was 2.48% (20/805), among which TypeⅠ(lacking CD36 expression both on platelet and monocyte) and TypeⅡ(lacking CD36 expression on platelet only) CD36 deficiency accounted for 10% (2/20) and 90% (18/20), respectively. CD36 gene mutations were found in 10 samples, including 3 cases of 329_330 delAC, 1 case of 1228_1239 delATTGTGCCTATT and 2 cases of 1163 A>T; 1 case of 329_330 delAC+ 1172_1183 delTATTGGTCAAGC and 287 G>C+ 329_330 delAC heterozygous mutation. In addition, 1 case of 745 A>G and 1 case of 806 C>T mutations were novel, and not yet reported. 【Conclusion】 Results showed that the frequency of CD36 antigen deficiency in Suzhou were similar to that reported in southern China, but the mutation sites were slightly different. The establishment of CD36 negative platelet registry could provide negative platelets for patients with transfusion reactions caused by anti-CD36 antibody and improve the effect of clinical platelet transfusion.
8.A CRISPR activation screen identifies genes that enhance SARS-CoV-2 infection.
Fei FENG ; Yunkai ZHU ; Yanlong MA ; Yuyan WANG ; Yin YU ; Xinran SUN ; Yuanlin SONG ; Zhugui SHAO ; Xinxin HUANG ; Ying LIAO ; Jingyun MA ; Yuping HE ; Mingyuan WANG ; Longhai TANG ; Yaowei HUANG ; Jincun ZHAO ; Qiang DING ; Youhua XIE ; Qiliang CAI ; Hui XIAO ; Chun LI ; Zhenghong YUAN ; Rong ZHANG
Protein & Cell 2023;14(1):64-68