Preparation of human monoclonal anti-C cell line from peripheral blood B lymphocytes of D--donor / 中国输血杂志

【Objective】 To establish human hybridoma cell lines, secreting monoclonal antibody against antigens of Rh blood system, from a donor with rare D--phenotype. 【Methods】 Peripheral blood B lymphocytes of an O type female donor, lacking C/c/E/e antigens on her erythrocyte, were transformed with Epstein-Barr virus (EBVs). EBVs were harvested from the cultural supernatant of B95-8 cells. The transformed lymphoblastoid cell line (LCL) secreting antibodies to C antigens were picked up and then hybridized with the myeloma SHM-D33 using electric fusion technique. Hybridoma cells were selected by HATD-Ouabain(HOTD)(Hypoxantine, Aminopterin, Thymidine, 2-Deoxycytide, and Ouabain)culture medium, microplate antibody screening and limited dilution subcloning. The monoclonal antibody was assayed by serological test and was confirmed by flow cytometry (FCM). 【Results】 From the cultural supernatant of D--peripheral blood transformed B lymphocytes, 3A6-C6, which agglutinated with R1R1(DCe/DCe)O-type RBCs but not with R2R2(DcE/DcE)O-type RBCs, was screened and preliminarily identified as anti-C. We established a hybridoma cell line secreting anti-C immunoglobulin M from B cells of D--individual successfully after hybridization with SHM-D33 myeloma cells. 【Conclusion】 The study had laid the groundwork for future research and development of human monoclonal antibodies against Rh antigens of RBC in future for diagnosis and screening purpose.

Risk factor analysis of plasma donation related vasovagal reaction / 中国输血杂志

【Objective】 To investigate the risk factors of vasovagal reactions(VVR) related to plasma donation, so as to put forward clinical suggestions for early identification and accurate intervention of high-risk groups to ensure the safety of plasma donation. 【Methods】 The demographic characteristics(i.e. gender, age, weight) and records of plasma donors(donation history, pulse before plasma donation, duration of collection, etc.) were collected from July to December 2019 in a region of Sichuan. Based on logistic regression analysis, the correlation between these factors and the risk of VVR was explored. 【Results】 The information of 69 172 donors was collected, and the incidence of VVR was 7.04‰. The risk of VVR was reduced by 99% in the group with plasma collection duration less than 30 minutes compared with the group with plasma collection duration more than 50 minutes(OR, 0.01; 95% CI, 0.00~0.01; P<0.001). The risk of male group was 94 % lower than that of female group(OR, 0.06; 95% CI, 0.04~0.10; P<0.001). Compared with the 45~50 kg group, the risk of weight greater than 80 kg group decreased by 80%(OR, 0.20; 95% CI, 0.09~0.42; P<0.001). The risk of repeated donation group was 34 % lower than that of the first time donation group(OR, 0.66; 95% CI, 0.47~0.91; P<0.001). The risk of VVR in the group with pulse greater than 90 bpm before plasma donation was 2.43 times that in the 60~69 bmp group(OR, 2.43; 95% CI, 1.75~3.36; P<0.001). 【Conclusion】 Duration of plasma collection, gender, weight, frequency of plasma donation, pulse before plasma donation and donor status are independent risk factors for plasma donation-related VVR. Among them, plasma collection duration, gender and weight were the main independent risk factors for plasma donation-related VVR. For donors with plasma collection duration more than 50 minutes, female and low weight, higher risk of VVR was presented and more preventive intervention should be given.

Characteristics of donors in platelet bank of Taiyuan and application evaluation of the bank / 中国输血杂志

【Objective】 To establish the donor bank for local region and distinguish the donors based on the past blood donation frequency and blood donation type, so as to improve the application efficiency of platelet bank. 【Methods】 1) According to the donation type and frequency of blood donors, the blood donors who had joined China Marrow Donor Program(CMDP)in our center from 2011 to 2020 were screened and classified. They are classified as reserve donors, prospective donors, and active donors. The donors, who met the selection conditions of active donors, were enrolled from all apheresis donors in 2020 to expand the local platelet bank. 2) In 2020, 739 blood donors who met the conditions of active donors were screened(including donors who had entered CMDP), and their HLA-A/B loci were detected by HLA high-resolution genotyping, and HPA was detected by Q-PCR genotyping. 3) The compatible platelets provided by three types of donors in 2021 were calculated. 【Results】 1) Taiyuan platelet bank, composed of donors with different previous donation experiences, had been constructed, including 739 active donors, 3840 prospective donors and 18 715 reserve donors. The composition ratio of ABO blood groups among three types of donors was found to be similar via chi square test; there were more male than female in three groups. 2) In 2021, the ratio of the average redonation by active donors in the platelet bank to the regular donation by the general donors was 14.4∶3.98. 3) Of the 142 compatible platelets, supplied to PTR patients in 2021, 83.8% of them came from the redonation of active donors after registration in the bank, and 9.9% and 6.3% from prospective donors and reserve donors, respectively. 4) In 2021, 28.1% of the stored pheresis platalets in our center had HLA typing data, and 54.2% of compatible platelets were retrieved from the inventory, which timely met the needs of clinical patients. 【Conclusion】 Integrating resources and distinguishing the activity degree of donors in platelet bank can reduce the cost of bank building and improve the application efficiency of the platelet bank.

Research progresses in donation related vasovagal reaction / 中国输血杂志

Donation related vasovagal reaction(DRVR) is the most common adverse reaction during blood donation. It is very important for blood banks to identify, treat and prevent DRVR accurately. At present, it is generally believed that psychological factors are the first major inducement of DRVR. Applied muscle tension (AMT) and salt supplementation have been proved to be effective interventions for vasovagal response; the identification methods of high-risk groups such as State Trait Anxiety Inventory, Medical Fear Inventory and Blood Donor Response Scale have been relatively mature, but the utilization rate is relatively low in China. In this paper, the main clinical manifestations, pathogenesis, research methods, related factors, management and prevention measures of DRVR, as well as the identification of high-risk groups before blood donation are reviewed.

-D- phenotype screening by polybrene method / 中国输血杂志

【Objective】 To develop a novel screening reagent for -D- phenotype preliminary screening based on the difference in RhD antigen expression level of -D- phenotype and normal RhD phenotype. 【Methods】 RhD antigen expression of -D-phenotype and Rh D-- gene carrier were detected by flow cytometry. By adjusting the concentration of polybrene in the screening system, the red blood cells with high RhD antigen expression level agglutinated, and the preliminary screening of the -D-phenotype and its gene carriers was realized. 【Results】 According to the quantitative results of immunofluorescence intensity (MFI) analysis by flow cytometry, the expression level of RhD antigen in -D- phenotype cells (284 360±16 698, n=3) was about 3 times normal RhD positive cells (98 642±35 908, n=9)(P<0.01), while RhD antigen expression level of RhD-- gene carrier (181 109±39 455, n=4) was about 2 times normal RhD positive cells(P<0.01). RhD antigen expression (144 538±227 445, n=7) of the positive cells screened by 15 μL 3% fresh red blood cell suspension and screening system 35 μL (1 μL IgG anti-D, 29 μL polybrene polybrene, and 5 μL low ionic strength solution) was about 1.5 times normal RhD positive cells. 【Conclusion】 The polybrene preliminary screening system, which can be used for high-throughput screening of -D- phenotype, is a reliable technical method for frequency study of this phenotype.

HLA-DRB1 genotyping in children with nephrotic syndrome of non-IgA mesangial proliferative giomerulonephritis / 中华微生物学和免疫学杂志

Objective To investigate the association of HLA-DRB1 alleles in Han population of Shanxi childrcn with nephrotic syndrome of non-IgA mesangial proliferative glomerulonephritis (MsPGN). Methods HLA-DRB1 was performed by polymerase chain reaction-sequence specific primers technique, and twenty patients with nephrotic syndrome of non-IgA MsPGN were detected. Results Analysis of the fre- quencies of specific at the HLA-DRB1 loci revealed significantly higher frequencies of HLA-DRB1 * 11 al- leles among the nephrotic syndrome patients of non-IgA MsPGN comparing with controls (22. 50% vs 8.33%, x2= 9. 544, P = 0.002, CI = 1. 674-9.995, RR = 4.09). Nine patients with HLA-DRB1 * 11 all accompanied hematuria, hypertension or short renal insufficiency. Conclusion The results suggested that HLA-DRB1 * 11 alleles contribute to genetic susceptibility to nephritic syndrome of non-IgA MsPGN. The pa- tients with HLA-DRB1 *11 easy accompanied hematuria, hypertension or short renal insufficiency.

Relationship between Host Survival and the Type of Immune Response in Different Organs during Disseminated Candidiasis / 华中科技大学学报（医学）（英德文版）

To examine the relationship between host survival and the type of immune response in different organs during disseminated candidiasis, the murine model of disseminated candidiasis was established by injection with Candida albicans via tail vein. The survival time was observed for up to 60 days. And the expression levels of cytokines in the spleen and kidney, including IFN-γ and IL-4, were determined with RT-PCR. Our results showed that in the spleen, both non-fatal and fatal inoculum caused a type Ⅱ immune response with steady expression levels of IFN-γ and the obviously increased levels of IL-4. While in the kidney, non-fatal inoculum induced a type Ⅰ immune response with the obviously increased levels of IFN-γ and the steady expression levels of IL-4. However, fatal inoculum induced a type Ⅱ immune response with a constant expression of IFN-γ and the evidently increased levels of IL-4. It is concluded that in disseminated candidiasis, host survival is associated with the type of immune responses in the kidney, but not in the spleen.

Susceptibility of mixed infection of Ureaplasma Urealyticum and Mycoplasma Hominis to seven antimicrobial agents and comparison with that of Ureaplasma Urealyticum infection / 华中科技大学学报（医学）（英德文版）

In order to investigate the susceptibility of mixed infection of Ureaplasma Urealyticum (UU) and Mycoplasma Hominis (MH) to 7 kinds of antimicrobial agents and comparison with that of UU infection in NGU patients, the in vitro susceptibility was determined by using microdilution method. The positive results were analyzed. The results showed that the sequence of susceptibility to 7 kinds of antimicrobial agents for both UU infection group and UU-MH mixed infection group was almost the same from the highest susceptibility to the lowest accordingly: Josamycin, Doxycycline, Minocycline, Sparfloxacin, Roxithromycin, Ofloxacin and Azithromycin. The total drug resistance rate for UU-MH mixed infection group (97.67%) was significantly higher than that for UU infection group (44.67%, P < 0.01). The highest drug resistance rate in UU group and UU-MH mixed infection group was 31.33% (Ofloxacin) and 90.48% (Azithromycin) respectively. UU-MH mixed infection showed an increased drug resistance and changes of drug resistance spectrum.

c Ha ras Proto oncogene and Psoriasis / 中华皮肤科杂志

The ras oncogene product ras p21, which is structurally homologous to G protein, participates in the regulation of cellular proliferation and differentiation. The histopathology of psoriasis is characterized by hyperkeratosis and parakeratosis. The purpose of this study was to investigate, by dot blot and Dig DNA labeling in situ hybridization techniques, the expression of c Ha ras proto oncogene in normal skin and active plaque psoriatic lesions. The results showed: (1) dot blot revealed one fold elevation of c Ha ras gene transcript levels in psoriatic lesions compared with normal controls. (2) In normal epidermis, only a few basal cells expressed ras gene, however, the ras gene was expressed throughout all layers of epidermis of active psoriatic lesions except horny layer and predominantly in prickle cells. Our results suggest that overexpression of c Ha ras oncogene and changes of expression pattern might be central to the pathogenesis of psoriatic epidermal hyperplasia and abnormal pattern of cellular differentiation.