1.A new method for flow cytometry-based detection of ABO antigen expression levels
Yuyu ZHANG ; Xi LIU ; Junhua XIE ; Bin CAO ; Jiewei ZHENG ; Xinyi ZHU ; Zhongying WANG ; Dong XIANG
Chinese Journal of Blood Transfusion 2025;38(5):665-672
Objective: To design and establish a new method for flow cytometry-based detection of commonly observed highly expressed antigens on red blood cells, and to further evaluate the differences and distribution characteristics of antigen expression levels between ABO blood type homozygotes and heterozygotes in healthy individuals. Methods: Residual blood samples after donor blood type identification by Shanghai Blood Center in April 2024 were collected. Among them, samples of 19 homozygous and 19 heterozygous individuals of type A and type B were selected. Then the expression level of ABO antigen on red blood cells were detected using the new method established in this study and the traditional aldehyde fixed red blood cell method. Both methods were tested independently three times and the results were compared. Results: The mean values of the three detection results of the new method was (×10
/RBC): AA homozygous 3.3±0.5, AO heterozygous 2.8±0.3, BB homozygous 3.6±0.3, BO heterozygous 3.1±2.8. The mean values of the three detection results of the aldehyde fixation method were AA homozygous 5.9±0.9, AO heterozygous 5.0±1.4, BB homozygous 3.8±0.6, and BO heterozygous 3.3±0.4. The average antigen distribution of each genotype followed a normal distribution. Comparing the average antigen expression levels of homozygotes and heterozygotes, both methods showed that A/B homozygotes had higher antigen levels than heterozygotes, with AA being 1.17 to 1.18 times that of AO and BB being 1.15 to 1.16 times that of BO. Comparing the inter batch differences in the three test results of two methods, the new method showed no significant difference in the three test results for four genotypes (P>0.05). The aldehyde fixation method showed significant differences in the test results for all three genotypes (P<0.01) except for BB homozygotes (P>0.05). The reliability and reproducibility of the new method were better than those of the traditional aldehyde fixation method. Conclusion: The antigen expression level of ABO homozygotes is higher than that of heterozygotes, and the difference in antigen level between type A homozygotes and heterozygotes is slightly higher than that of type B. The new method is superior to traditional aldolization fixation methods.
2.Hemolytic disease of the fetus and newborn caused by Rh system anti-c antibodies: a case report and literature review
Luyan CHEN ; Dong XIANG ; Dingfeng LYU ; Zhenyun LIU ; Xinyi ZHU ; Shuan TAO ; Qiming YING ; Wei LIANG
Chinese Journal of Blood Transfusion 2025;38(6):843-848
Objective: To summarize the laboratory findings of a case of hemolytic disease of the fetus and newborn (HDFN) caused by Rh system anti-c antibodies and to review the literature, so as to explore the characteristics of anti-c HDFN. Methods: The ABO blood type, Rh blood type, direct antiglobulin test (DAT) results, and the presence of unexpected antibodies and their titers were determined by serological methods. The cases of anti-c HDFN in our laboratory in China and abroad were statistically analyzed, and the incidence of severe HDFN caused by anti-c, anti-D and anti-E was compared. Results: The blood type of the child was B (Rh CcDee) with a positive DAT. Anti-c antibody was detected in both serum and eluate, with a serum antibody titer of 4. The mother’s blood type was AB (Rh CCDee) with a negative DAT, and anti-c antibody was detected in the serum with a titer of 128. Among 20 cases of anti-c HDFN, 17 were DAT positive, and 9 (45%, 9/20) underwent blood transfusion or exchange transfusion. The incidence of severe HDFN was 47.60% (10/21) for anti-c, 47.60% (10/21) for anti-D and 31.30% (5/16) for anti-E. Conclusion: Maternal pregnancy and/or blood transfusion are the main reasons for the production of Rh alloantibodies such as anti-c. The prevention and management of anti-c should be similar to that of anti-D. Rh antigen-matched (five antigens of Rh blood group) transfusion is necessary for women of childbearing age to avoid antibody production, and Rh typing and antibody screening during prenatal examination is recommended to ensure early detection, intervention and treatment.
3.Opportunities and challenges of marginal donor liver
Xinyi LU ; Fei TENG ; Hong FU ; Yuanyu ZHAO ; Liye ZHU ; Jiayong DONG ; Jiaxi MAO ; Wenyuan GUO
Organ Transplantation 2024;15(3):463-468
With persistent breakthrough and maturity of surgical procedures and postoperative immunosuppressive therapy, the survival rate of liver transplant recipients and grafts has been significantly increased. The shortage of donor liver has become the main obstacle for clinical development of liver transplantation. How to expand the source of donor liver has become an urgent issue. Groundbreaking progresses have been made in the use of common marginal donor livers in clinical liver transplantation, such as elderly donor liver, steatosis donor liver, viral hepatitis donor liver and liver from donation after cardiac death. Nevertheless, multiple restrictions still exist regarding the use of marginal donor liver. Consequently, the definition of marginal donor liver and research progress in the application of common marginal donor livers were reviewed, and the opportunities and challenges of mariginal donoor liver were illustrated, aiming to provide reference for expanding the donor pool for clinical liver transplantation and bringing benefits to more patients with end-stage liver disease.
4.Application of Ancient Books in Clinical Practice Guidelines and Expert Consensus of Traditional Chinese Medicine: Current Status and Methodological Recommendations
Changhao LIANG ; Dingran YIN ; Jing CUI ; Xinshuai YAO ; Xinyi GU ; Yifei YAN ; Wanting LIU ; Yingqiao WANG ; Yingqi CHANG ; Haoyu DONG ; Mengqi LI ; Yuanyuan LI ; Yutong FEI
Journal of Traditional Chinese Medicine 2024;65(8):801-809
ObjectiveTo explore the current status and issues regarding the application of ancient books in clinical practice guidelines and expert consensus of traditional Chinese medicine (TCM) published in China, and to provide methodological recommendations for the incorporation of ancient books in the development of TCM guidelines. MethodsWe searched China National Knowledge Infrastructure (CNKI), WanFang Data, VIP, SinoMed, PubMed, Embase, as well as six industry websites including China Association of Chinese Medicine, National Group Standards Information Platform, and Chinese Association of the Integration of Traditional and Western Medicine,etc. TCM clinical practice guidelines or expert consensus issued during January 1st, 2017, to November 26th, 2022 were searched. Clinical practice guidelines or expert consensus that explicitly referred to ancient books were included, and the content regarding the searching for ancient books, sources of access to ancient books, methods of evaluating the level of evidence, methods of evaluating the level of recommendation, and methods of evaluating the evidence for the ancient books were analysed. ResultsA total of 1,215 TCM clinical practice guidelines or expert consensus were retrieved, with 442 articles explicitly mentioning the application of ancient books, including 300 (67.87%) clinical practice guidelines and 142 (32.13%) expert consensus. Sixty of the 442 publications explicitly reported that ancient books searching had been conducted (13.57%); among these 60 publications 27 (45.00%) explicitly reported ancient books searching strategies, and the most frequent method was manual searching with a total of 24 articles (40.00%). The most popular search source was Chinese Medical Dictionary, a TCM classics database, with a total of 18 articles. 197 articles (44.57%) explicitly reported the evaluation criteria for the level of evidence, of which 141 articles (71.57%) involved the evaluation criteria for the ancient books; 413 articles (93.44%) mentioned ancient books in the recommendations, and only the source of formula name was mentioned in 409 (99.03%) of the publications. ConclusionThe current application of ancient books in TCM clinical practice guidelines and expert consensus is limited, with issues of non-standard searching and evaluation methods. Standar-dization and uniformity are needed in evidence grading and recommendation standards. Future research should clarify the scope and methods of applying ancient book, emphasize their integration with modern research evidence, and enhance their value and quality in the development of TCM clinical practice guidelines.
5.Quantitative research on China's disability rehabilitation policy using policy modeling consistency index model
Tongtong GUO ; Xinyi ZHANG ; Lihong JI ; Zhiwei DONG ; Zongrun LI ; Liduan WANG ; Weiqin CAI ; Qianqian GAO ; Qi JING ; Wengui ZHENG
Chinese Journal of Rehabilitation Theory and Practice 2024;30(6):621-629
Objective To quantitatively analyze and evaluate the content of rehabilitation policy for people with disabilities in China. Methods This study focused on ten national policies of disability rehabilitation issued from 2021 to 2023.It employed text mining techniques to process policy texts and constructed a policy modeling consistency index model for dis-ability rehabilitation policies in China.The relevant policies were evaluated and analyzed quantitatively. Results The disability rehabilitation policies in China were relatively comprehensive in terms of policy transparency,op-erational mechanisms and policy nature.However,there was still a need for optimization in terms of policy per-spectives,target groups,incentive mechanisms,and other aspects. Conclusion The overall quality of disability rehabilitation policy texts at the national level in China is relatively good.There is a need to further enhance the predictability of policy objectives,clarify the responsibilities and division of labor among various departments,and improve policy incentive mechanisms in future policy formulation,which will further refine China's disability rehabilitation policy system and contribute to high-quality develop-ment of the disability cause.
6.Clinical Efficacy of Erdong Xiaoke Formula in Treating Type 2 Diabetes Dry Eyes with Yin Deficiency and Heat Excess Syn-drome and Its Influence on Serum IL-17 and IL-1β
Xinyi SUN ; Huijie JI ; Li SHI ; Huxing SHEN ; Jiangyi YU ; Yingying DONG
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(3):302-308
OBJECTIVE To observe the clinical efficacy of Erdong Xiaoke Formula in the treatment of type 2 diabetes dry eyes with yin deficiency and heat excess syndrome and its effect on serum interleukin 17(IL-17)and interleukin 1β(IL-1β),and to ex-plore the therapeutic mechanism of Erdong Xiaoke Formula.METHODS 110 cases of type 2 diabetes patients with dry eyes of yin deficiency and heat excess syndrome from Jiangsu Province Hospital of Chinese Medicine were enrolled and randomly divided into a treatment group and a control group,55 cases each.5 cases dropped out of the treatment group and 4 cases dropped out of the control group.The control group was given a basic hypoglycemic regimen combined with topical sodium hyaluronate eye drops.The treatment group was given Erdong Xiaoke Formula in addition to the treatment in the control group.The treatment course for both groups was 4 weeks.Changes in TCM syndrome scores of the two groups of patients were observed before and after treatment,and the clinical effica-cy of TCM was evaluated.Blood glucose and pancreatic islet function-related indicators[fasting blood glucose(FBG),fasting insulin(FINS),fasting C-peptide(FCP),postprandial 2 h blood glucose(PBG),insulin secretion function index(HOMA-β),insulin re-sistance index(HOMA)-IR)],ocular surface indicators[tear break up time(BUT),corneal sodium fluorescein staining(FL),Schirmer Ⅰ test(SⅠT)],and inflammation-related indicators(IL-17,IL-1β)were detected.The occurrence of adverse reactions in the two groups of patients was observed during the treatment period.RESULTS After treatment,the total scores of TCM syn-dromes in both groups were significantly reduced(P<0.01),the treatment group was better than the control group(P<0.01),and the total effective rate of TCM syndromes in the treatment group was higher than that of the control group(P<0.01);SⅠT and BUT of pa-tients in both groups increased significantly(P<0.01),and the treatment group was better than the control group(P<0.05,P<0.01);the FL of patients in both groups significantly reduced(P<0.01),and there was no significant difference between the groups;the ser-um IL-17 and IL-1β of the patients in the treatment group decreased significantly(P<0.01),which was significantly better than that of the control group(P<0.05).There were no significant changes in blood glucose and pancreatic islet function-related indicators in the two groups before and after treatment(P>0.05).During the treatment,no obvious adverse reactions were observed in the two groups.CONCLUSION Erdong Xiaoke Formula can improve SⅠT,BUT,FL and eye symptoms in patients with diabetic dry eye,effectively treat diabetic dry eye,and reduce ocular surface inflammation.Its mechanism may be related to reducing serum inflammato-ry factors IL-17 and IL-1β.
7.Molecular mechanism of lncRNA SNHG1 regulating ferroptosis and at-tenuating inflammation of microglia induced by HIV-1 gp120 V3 loop
Linlin WANG ; Qin ZUO ; Xinyi LI ; Xueqin YAN ; Rui PAN ; Yongmei FU ; Jun DONG
Chinese Journal of Pathophysiology 2024;40(5):806-814
AIM:To investigate the molecular mechanism of long noncoding RNA(lncRNA)SNHG1 in regu-lating ferroptosis to alleviate inflammation in CHME-5 human microglia induced by HIV-1 gp120 V3 loop.METHODS:CHME-5 human microglia were cultured in vitro,and were divided into 7 groups:blank group,random peptide group,gp120 V3 loop group(HIV-1 gp120 group),HIV-1 gp120+shCon group,HIV-1 gp120+SNHG1 sh2 group,HIV-1 gp120+SNHG1 sh2+ferrostatin-1(Fer-1;ferroptosis inhibitor)group,and HIV-1 gp120+SNHG1 sh2+EX527(Sirt1 in-hibitor)group.Normal CHME-5 cells were treated with random peptide or gp120 V3 loop for 24 h.After pretreatment of SNHG1 sh2 cells with inhibitors for 2 h,the cells were then treated with gp120 V3 loop for 24 h.The levels of inflammato-ry cytokines in the cell supernatants were detected by ELISA.Western blot was used to detect the protein expression levels of solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),Sirt1 and p53.Microplate reader was used to detect the levels of intracellular ferrous ion(Fe2+)and malondialdehyde(MDA).RESULTS:(1)The results of ELISA showed that the expression levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and IL-1β in HIV-1 gp120 group were significantly higher than those in blank group(P<0.05).Compared with HIV-1 gp120 group,the ex-pression levels of inflammatory cytokines in HIV-1 gp120+SNHG1 sh2 group were significantly decreased(P<0.05).Compared with HIV-1 gp120+SNHG1 sh2 group,the expression levels of inflammatory factors in HIV-1 gp120+SNHG1 sh2+Fer-1 were significantly decreased(P<0.05),but those in HIV-1 gp120+SNHG1 sh2+EX527 group were significant-ly increased(P<0.01).(2)The results of Western blot showed that compared with blank group,the expression levels of ferroptosis-related proteins SLC7A11 and GPX4 in HIV-1 gp120 group were significantly down-regulated(P<0.01).Com-pared with HIV-1 gp120 group,the expression levels of SLC7A11 and GPX4 in HIV-1 gp120+SNHG1 sh2 group were sig-nificantly up-regulated(P<0.01).Compared with HIV-1 gp120+SNHG1 sh2 group,the expression levels of SLC7A11 and GPX4 in HIV-1 gp120+SNHG1 sh2+Fer-1 group were significantly up-regulated(P<0.05),but the expression levels of SLC7A11 and GPX4 in HIV-1 gp120+SNHG1 sh2+EX527 group were significantly down-regulated(P<0.01),and the expression level of p53 was significantly up-regulated(P<0.05).(3)Compared with blank group,the levels of Fe2+and MDA in HIV-1 gp120 group were significantly increased(P<0.05).Compared with HIV-1 gp120 group,the levels of Fe2+and MDA in HIV-1 gp120+SNHG1 sh2 group were significantly decreased(P<0.01).Compared with HIV-1 gp120+SNHG1 sh2 group,Fe2+and MDA in HIV-1 gp120+SNHG1 sh2+Fer-1 group were significantly decreased(P<0.05),but those in HIV-1 gp120+SNHG1 sh2+EX527 group was significantly increased(P<0.05).CONCLUSION:Knockdown of SNHG1 can attenuate the inflammation in microglia induced by HIV-1 gp120 V3 loop,which may be achieved by regulating ferrop-tosis-related signaling molecules through the Sirt1/p53 signaling pathway.
8.Molecular characteristics and drug susceptibility analysis of Streptococcus agalactiae from respiratory specimen sources
Xiao HAN ; Xinyi GONG ; Beibei MIAO ; Huan XING ; Zeliang LIU ; Pengfang GAO ; Yuelong LI ; Jiachen LI ; Yating TANG ; Yanlei GE ; Aiying DONG ; Juan LI
Chinese Journal of Preventive Medicine 2024;58(6):891-897
To study the carriage status of drug susceptibility, clonal complex groups, serotypes, surface proteins and virulence genes of Streptococcus agalactiae from respiratory specimen sources. A total of 35 strains of S.agalactiae meeting the criteria were collected from 3 hospitals in 2 locations, Tangshan and Jinan. The age span of the patients was 3 days-92 years, and the percentage of elderly patients≥60 years was 71.5%.The susceptibility to 9 antimicrobial drugs was measured and analyzed using the micro broth dilution method. The strains were 100.0% sensitive to penicillin, linezolid, vancomycin, and ceftriaxone; However, it exhibits high resistance rates to erythromycin, clindamycin and levofloxacin, at 97.1%, 85.7% and 82.9% respectively; and the resistance rates to tetracycline and chloramphenicol were 34.3% and 14.2%, respectively. Genome sequence determination and analysis showed that 16 resistance genes were detected in 35 strains, among which: macrolide and lincosamide resistance genes were mainly ermB, with a carrying rate of 74.2%; tetracycline resistance genes were mainly tetM, with a carrying rate of 25.7%; in addition, the mutation rates of the quinolone resistance determinants gyrA and parC were 88.5% and 85.7%, respectively. 35 strains belonged to 6 ST types and 4 clonal groups, with CC10/ST10 as the main one, accounting for 62.8%; they contained 4 serotypes of Ⅰb, Ⅱ, Ⅲ, and Ⅴ, as well as 1 untyped strain, with serotype Ⅰb as the main one, accounting for 65.7%. The strains carried three pilus types, PI1+PI2a, PI2a and PI2b types, respectively, and detected five surface proteins, alpha, alp1, rib, srr, and r df_0594, and seven virulence factors, cba, cfb, cylE, fbsA, hylB, l mb, and pavA. Overall, S.agalactiae isolated from respiratory tract specimens is predominantly sourced from elderly patients, with CC10 strains being most prevalent. These strains harbor multiple drug-resistant and virulence genes, demonstrating elevated resistance rates to macrolides, lincosamides, and quinolones. This emphasizes the necessity for vigilant attention to the health threat posed by S. agalactiae from respiratory tract speciments of elderly patients.
9.TCM-HIN2Vec: A strategy for uncovering biological basis of heart qi deficiency pattern based on network embedding and transcriptomic experiment
Lihong Diao ; Xinyi Fan ; JIang Yu ; Kai Huang ; Edouard C Nice ; Chao Liu ; Dong Li ; Shuzhen Guo
Journal of Traditional Chinese Medical Sciences 2024;11(3):264-274
Objective:
To elucidate the biological basis of the heart qi deficiency (HQD) pattern, an in-depth understanding of which is essential for improving clinical herbal therapy.
Methods:
We predicted and characterized HQD pattern genes using the new strategy, TCM-HIN2Vec, which involves heterogeneous network embedding and transcriptomic experiments. First, a heterogeneous network of traditional Chinese medicine (TCM) patterns was constructed using public databases. Next, we predicted HQD pattern genes using a heterogeneous network-embedding algorithm. We then analyzed the functional characteristics of HQD pattern genes using gene enrichment analysis and examined gene expression levels using RNA-seq. Finally, we identified TCM herbs that demonstrated enriched interactions with HQD pattern genes via herbal enrichment analysis.
Results:
Our TCM-HIN2Vec strategy revealed that candidate genes associated with HQD pattern were significantly enriched in energy metabolism, signal transduction pathways, and immune processes. Moreover, we found that these candidate genes were significantly differentially expressed in the transcriptional profile of mice model with heart failure with a qi deficiency pattern. Furthermore, herbal enrichment analysis identified TCM herbs that demonstrated enriched interactions with the top 10 candidate genes and could potentially serve as drug candidates for treating HQD.
Conclusion
Our results suggested that TCM-HIN2Vec is capable of not only accurately identifying HQD pattern genes, but also deciphering the basis of HQD pattern. Furthermore our finding indicated that TCM-HIN2Vec may be further expanded to develop other patterns, leading to a new approach aimed at elucidating general TCM patterns and developing precision medicine.
10.Impact of embryonic uveitis exposure on response of mouse offspring to interphotoreceptor retinoid-binding protein-induced experimental autoimmune uveitis
Fei XU ; Jianping LIU ; Shifang DONG ; Hui HUANG ; Xinyi GONG ; Kaijiao HU ; Feilan CHEN
Acta Laboratorium Animalis Scientia Sinica 2024;32(3):297-306
Objective To investigate the effect of embryonic inflammatory exposure on the response of mouse offspring to interphotoreceptor retinoid-binding protein(IRBP)-induced experimental autoimmune uveitis(EAU).Methods RNA transcriptome sequencing data from eyeballs of C57BL/6J mouse offspring born to mothers with active EAU were used to screen immune-associated differentially expressed genes in the eyes of the exposed offspring.Gene fragments overlapping in the two datasets were screened using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses to identify biological pathways associated with the gene fragments.Hub genes were identified from these intersecting genes by protein-protein interaction network analysis.EAU models of maternal uveitis were established by immunization with IRBP651-670,and expression levels of the pivotal genes in the offspring exposed to inflammation by maternal uveitis were examined by fluorescence quantitative polymerase chain reaction.EAU severity,T lymphocyte proliferation,and serum cytokines were detected to investigate the immune effect in offspring from mothers with an active inflammation response to IRBP induction.Results Microarray analysis identified 72 immune-related differentially expressed genes in exposed samples compared with the findings in control samples.These genes were mainly enriched in Toll-like receptor signaling,mitogen-activated protein kinase signaling,and B cell receptor signaling pathways.Protein-protein interaction network interaction analysis screened out four hub genes,Psmc5,Psmc3,Psmd4,and Psmd8,and mRNA levels of these four genes were increased in the adult offspring from mothers with active uveitis compared with the findings in healthy offspring.In addition,the group induced with 150 μg IRBP showed an increase in the severity of clinical and pathological outcomes in offspring with EAU affected by active inflammation,compared with the healthy offspring group(P=0.0087,P=0.0410).Meanwhile,T cell proliferation in the offspring was enhanced during the inflammatory activity stage and secretion of the inflammatory cytokines interleukin(IL)-17 and IL-6 was increased(P=0.0450,P=0.0300).Conclusions Psmc5,Psmc3,Psmd4,and Psmd8 may be important genes exacerbating uveitis in offspring of mothers with active uveitis,associated with increased T cell proliferation and production of IL-17 and IL-6.


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