1.Application of serological methods combined with flow cytometry in the detection of immune hemolytic transfusion reaction
Lei LIU ; Qunjuan ZENG ; Guoqin GONG ; Dong XIANG ; Zhongying WANG
Chinese Journal of Blood Transfusion 2025;38(1):116-121
[Objective] To explore the effects of different methods on antibody detection through investigating the causes of cross-matching incompatible in a patient with gastric malignant tumor, and to establish flow cytometry protocol for confirming hemolytic transfusion reaction (HTR). [Methods] Antibodies in the patient's serum were identified by red blood cells (RBCs) blood grouping, antibody screening and identification, acid elution test and PEG enhancement test. To confirm HTR, patient RBCs, proximal and distal ends RBCs, separated by capillary centrifugation, were tested by direct antiglobulin test (DAT) and Jka antigen single label and double label flow cytometry. [Results] Routine serological technology revealed the presence of anti-C, e (titer:2) and anti-Jka (titer >1) in the patient’s serum. After separation using capillary centrifugation technology, both the proximal and distal DAT and Jka antigen tests were negative. Both DAT and Jka antigen positive red blood cells (0.21%, 6/6 327) were found in the patient's blood samples by flow cytometry. After separation of blood samples by capillary centrifugation, there were significantly more DAT and Jka antigen double-positive RBCs in the distal end (0.43%, 33/7 707) than in the proximal end (0.09%, 15/7 225). Two blood samples were screened from over 100 donor blood samples that are compatible with the patient's cross-matching, and the transfusion effect was favorable. [Conclusion] Serological methods combined with flow cytometry could improve the sensitivity of antibody detection, provide a more accurate basis for the diagnosis of HTRs, and guarantee the safety of blood transfusion.
2.Comparison of the efficacy of heat and acid elution methods for IgG anti-M and anti-Ku
Qunjuan ZENG ; Huaiying KANG ; Dong XIANG ; Wei SHEN ; Chengrui QIAN ; Zhongying WANG ; Guoqin GONG
Chinese Journal of Blood Transfusion 2025;38(7):964-968
Objective: To compare the efficacy of heat and acid elution methods for IgG anti-M and anti-Ku. Methods: Ten samples with IgG anti-M and two samples with IgG anti-Ku were selected and standardized to a titer of 64. These antibodies underwent overnight absorption at 4℃ with O-type MM and kk-type erythrocytes, and then heat and acid elution methods were used on the absorbed sensitized erythrocytes respectively by detecting the titer of anti-M and anti-Ku in the eluate to compare the differences in the elution efficiency of IgG anti-M and anti-Ku between the two elution methods. Results: In heat elution tests, all 10 anti-M samples showed positive results with titers ranging from 8 to 64, while 2 anti-Ku samples yielded negative results. In acid elution tests, all 10 anti-M samples demonstrated negative results, whereas both anti-Ku (n=2) samples exhibited positive reactions with consistent titers of 32. Following acid elution with subsequent heat elution, 8 of 10 anti-M samples showed positive results with titers ranging from 8 to 32, while 2 remained negative. Both anti-Ku samples demonstrated positive with titers of 4. Conclusion: Heat elution demonstrated superior efficiency for IgG anti-M compared to acid elution, whereas acid elution showed greater efficacy for IgG anti-Ku than heat elution.
3.Comparison of the efficacy of heat and acid elution methods for IgG anti-M and anti-Ku
Qunjuan ZENG ; Huaiying KANG ; Dong XIANG ; Wei SHEN ; Chengrui QIAN ; Zhongying WANG ; Guoqin GONG
Chinese Journal of Blood Transfusion 2025;38(7):964-968
Objective: To compare the efficacy of heat and acid elution methods for IgG anti-M and anti-Ku. Methods: Ten samples with IgG anti-M and two samples with IgG anti-Ku were selected and standardized to a titer of 64. These antibodies underwent overnight absorption at 4℃ with O-type MM and kk-type erythrocytes, and then heat and acid elution methods were used on the absorbed sensitized erythrocytes respectively by detecting the titer of anti-M and anti-Ku in the eluate to compare the differences in the elution efficiency of IgG anti-M and anti-Ku between the two elution methods. Results: In heat elution tests, all 10 anti-M samples showed positive results with titers ranging from 8 to 64, while 2 anti-Ku samples yielded negative results. In acid elution tests, all 10 anti-M samples demonstrated negative results, whereas both anti-Ku (n=2) samples exhibited positive reactions with consistent titers of 32. Following acid elution with subsequent heat elution, 8 of 10 anti-M samples showed positive results with titers ranging from 8 to 32, while 2 remained negative. Both anti-Ku samples demonstrated positive with titers of 4. Conclusion: Heat elution demonstrated superior efficiency for IgG anti-M compared to acid elution, whereas acid elution showed greater efficacy for IgG anti-Ku than heat elution.
4.Preparation and in vivo Distribution of Essential Oil from Alpinia zerumbet Fructus Encapsulated Nanoemulsions
Jinzhuan XU ; Lili ZHANG ; Zhengli ZHOU ; Shan XU ; Xiang ZHOU ; Lei HUANG ; Zipeng GONG ; Yi CHEN ; Xiangchun SHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):126-133
ObjectiveTo prepare oral nanoemulsions encapsulating essential oil from Alpinia zerumbet fructus(EOFAZ) and to investigate its pro-absorption effect in vitro and distribution in vivo. MethodThe proteoglycan conjugate polysaccharides of vinegar-processed Bupleuri Radix-bovine serum albumin(VBCP-BSA) was prepared by Maillard reaction of VBCP and BSA. Taking VBCP-BSA as emulsifier, vitamin B12(VB12) as absorption enhancer, and medium chain triglycerides mixed with EOFAZ as oil phase, the nanoemulsions loaded with EOFAZ was prepared by high energy emulsification method. The particle size, particle size distribution, surface Zeta potential, EOFAZ content and appearance and morphology of the nanoemulsions were characterized, and fluorescein tracer method was used to investigate the absorption effect of fluorescein-labeled EOFAZ nanoemulsions in vitro and their distribution in vivo. ResultVBCP-BSA was formed by Maillard reaction for 48 h with high grafting rate. Using VBCP-BSA as emulsifier, the homogeneous pink nanoemulsions was prepared and denoted as EOFAZ@VBCP-BSA/VB12. The particle size of the nanoemulsions was less than 100 nm and the particle size distribution was uniform. The surface of the nanoemulsions was a weak negative charge, and the shape was spherical. The encapsulation rate of the nanoemulsions for EOFAZ was greater than 80%, which had a good absorption effect in vitro and could enhance liver accumulation after oral administration. ConclusionThe designed proteoglycan nanoemulsions can effectively load EOFAZ, promote oral absorption and enhance liver distribution, which can provide experimental basis for the development of oral EOFAZ liver protection preparations.
5.Structure-based development of potent and selective type-II kinase inhibitors of RIPK1.
Ying QIN ; Dekang LI ; Chunting QI ; Huaijiang XIANG ; Huyan MENG ; Jingli LIU ; Shaoqing ZHOU ; Xinyu GONG ; Ying LI ; Guifang XU ; Rui ZU ; Hang XIE ; Yechun XU ; Gang XU ; Zheng ZHANG ; Shi CHEN ; Lifeng PAN ; Ying LI ; Li TAN
Acta Pharmaceutica Sinica B 2024;14(1):319-334
Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) functions as a key regulator in inflammation and cell death and is involved in mediating a variety of inflammatory or degenerative diseases. A number of allosteric RIPK1 inhibitors (RIPK1i) have been developed, and some of them have already advanced into clinical evaluation. Recently, selective RIPK1i that interact with both the allosteric pocket and the ATP-binding site of RIPK1 have started to emerge. Here, we report the rational development of a new series of type-II RIPK1i based on the rediscovery of a reported but mechanistically atypical RIPK3i. We also describe the structure-guided lead optimization of a potent, selective, and orally bioavailable RIPK1i, 62, which exhibits extraordinary efficacies in mouse models of acute or chronic inflammatory diseases. Collectively, 62 provides a useful tool for evaluating RIPK1 in animal disease models and a promising lead for further drug development.
6.Self-sufficient nanoparticles with dual-enzyme activity trigger radical storms and activate cascade-amplified antitumor immunologic responses.
Liping BAI ; Jin YANG ; Siting YU ; Zhongzheng XIANG ; Yuanyuan ZENG ; Meiling SHEN ; Xiaorong KOU ; Qinjie WU ; Changyang GONG
Acta Pharmaceutica Sinica B 2024;14(2):821-835
Radiotherapy (RT) can potentially induce systemic immune responses by initiating immunogenic cell death (ICD) of tumor cells. However, RT-induced antitumor immunologic responses are sporadic and insufficient against cancer metastases. Herein, we construct multifunctional self-sufficient nanoparticles (MARS) with dual-enzyme activity (GOx and peroxidase-like) to trigger radical storms and activate the cascade-amplified systemic immune responses to suppress both local tumors and metastatic relapse. In addition to limiting the Warburg effect to actualize starvation therapy, MARS catalyzes glucose to produce hydrogen peroxide (H2O2), which is then used in the Cu+-mediated Fenton-like reaction and RT sensitization. RT and chemodynamic therapy produce reactive oxygen species in the form of radical storms, which have a robust ICD impact on mobilizing the immune system. Thus, when MARS is combined with RT, potent systemic antitumor immunity can be generated by activating antigen-presenting cells, promoting dendritic cells maturation, increasing the infiltration of cytotoxic T lymphocytes, and reprogramming the immunosuppressive tumor microenvironment. Furthermore, the synergistic therapy of RT and MARS effectively suppresses local tumor growth, increases mouse longevity, and results in a 90% reduction in lung metastasis and postoperative recurrence. Overall, we provide a viable approach to treating cancer by inducing radical storms and activating cascade-amplified systemic immunity.
7.Effect of modified Baduanjin exercise on cardiopulmonary function, motor function and activities of daily living for stroke patients
Junwen CHEN ; Qian CHEN ; Cheng CHEN ; Shuyue LI ; Lingling LIU ; Cunshu WU ; Xiang GONG ; Jun LU ; Guangxu XU
Chinese Journal of Rehabilitation Theory and Practice 2024;30(1):74-80
ObjectiveTo investigate the effect of modified Baduanjin exercise, as an rehabilitation exercise, on cardiopulmonary function, motor function and activities of daily living in patients with stroke. MethodsFrom January to September, 2023, 42 stroke patients in the Nanjing Qixia District Hospital were randomly divided into control group (n = 21) and experimental group (n = 21). The control group received routine rehabilitation, and the experimental group received modified Baduanjin exercise in addition, for four weeks. They were assessed with peak oxygen uptake (VO2peak), anaerobic threshold (AT), peak oxygen pulse (VO2peak/HR), forced vital capacity (FVC), forced expiratory volume in the first second (FEV1), peak expiratory flow (PEF), Fugl-Meyer Assessment-upper extremities (FMA-UE), Berg Balance Scale (BBS) and modified Barthel Index (MBI) before and after intervention. ResultsVO2peak, AT, and the scores of FMA-UE, BBS and MBI improved in the control group after intervention (|t| > 2.256, |Z| > 2.936, P < 0.05); while VO2peak, AT, VO2peak/HR, FVC, FEV1, PEF, and the scores of FMA-UE, BBS and MBI improved in the experimental group (|t| > 4.390, |Z| > 3.451, P < 0.001); and all the indexes were better in the experimental group than in the control group (|t| > 4.136,|Z| > 2.751,P < 0.01), except the scores of BBS and MBI. ConclusionModified Baduanjin exercise can improve the cardiopulmonary function and upper limb motor function for stroke patients.
8.Role of Macrophage Activation and Polarization in Myocardial Fibrosis and Intervention of Traditional Chinese Medicine
Kunpeng YAO ; Huzhi CAI ; Xiang ZHAO ; Ke GONG ; Chuning TIAN ; Yuntao LUO ; Liqi PENG ; Guangyang OU ; Qingyang CHEN ; Xinyu CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):272-282
Myocardial fibrosis (MF) is a common pathological manifestation of various heart diseases. Due to the non-renewable nature of myocardial cells, the occurrence of MF represents irreversible damage to the myocardium. Previous studies have suggested that fibroblast-mediated collagen deposition is the main mechanism of MF. Recent studies have found that there is an immune regulation mechanism in the heart itself, and macrophage activation/polarization plays an important role in MF. With the deepening of traditional Chinese medicine research, scholars have found that traditional Chinese medicine can interfere with MF by regulating the renin-angiotensin-aldosterone system (RAAS) system and the inflammatory process, repairing the extracellular matrix, managing oxidative stress, and maintaining the balance of autophagy. This process is closely related to the activation and M1/M2 polarization of macrophages. Throughout the MF process, macrophage activation is beneficial, but excessive activation will be harmful. In the early stage of MF, appropriate M1 macrophage polarization is conducive to activating immunity and removing harmful substances. In the middle and late stages of MF, appropriate M2 macrophage polarization is conducive to remodeling the damaged myocardium. If macrophage activation is excessive/insufficient, or the balance of M1/M2 macrophage polarization is broken, the effect changes from improvement to destruction. Traditional Chinese medicines that regulate the activation/polarization of macrophages have the effects of replenishing Qi and nourishing Yin, as well as regulating Qi and activating blood, but there are also some heat-clearing, dampness-drying, and detoxification products. Therefore, the occurrence of MF may be caused by Qi and Yin deficiency, damp heat accumulation, and Qi stagnation and blood stasis. By summarizing the biological processes involved in macrophage activation/polarization in MF, this paper expounded on the research progress of traditional Chinese medicine in regulating macrophage activation and M1/M2 polarization from different angles to improve MF, so as to provide a reference for the treatment of MF with traditional Chinese medicine.
9.Effect of galvanic vestibular stimulation on stroke patients with lateropulsion
Xiang GONG ; Menghuan WANG ; Cunshu WU ; Junwen CHEN ; Yue XIAO ; Yun YANG ; Wanting SUN ; Jun LU ; Guangxu XU
Chinese Journal of Rehabilitation Theory and Practice 2024;30(2):210-216
ObjectiveTo investigate the effect of galvanic vestibular stimulation on stroke patients with lateropulsion. MethodsFrom February to October, 2023, 30 stroke patients with lateropulsion in the First Affiliated Hospital of Nanjing Medical University were divided into control group (n = 15) and experimental group (n = 15) randomly. The control group received routine rehabilitation and sham galvanic vestibular stimulation, and the experimental group received routine rehabilitation and true galvanic vestibular stimulation, for two weeks. They were assessed with Scale for Contraversive Pushing (SCP), subjective visual vertical (SVV), Line Cancellation Test (LCT), Star Cancellation Test (SCT), Berg Balance Scale (BBS), Holden Functional Ambulation Category (FAC) and Barthel Index (BI) before and after treatment. ResultsAll the indexes improved in both groups after intervention (|t| > 2.461, Z > 3.000, P < 0.05), except the SVV orientation, SVV certainty and SCT in the control group; while the SCP, SVV certainty, LCT and FAC were better in the experimental group than in the control group (|t| > 2.189, Z = -2.862, P < 0.05), and the differences before and after intervention were better in the experimental group than in the control group (|t| > 2.382, P < 0.05), except LCT. SCP was correlated with SVV orientation, SVV certainty, SCT, BBS, BI and FAC (|r| > 0.381, P < 0.05). ConclusionGalvanic vestibular stimulation can improve the lateropulsion, balance, walking function and activities of daily living in stroke patients, which may be related to improvement for spatial cognitive function, especially vertical perception.
10.Discussion on the pathogenesis of colorectal polyp from "damp pathogen causing diseases"
Zikuan SUN ; Xiaolan SU ; Yanran BAN ; Zhuozhi GONG ; Kaixuan ZHANG ; Lang XIANG ; Mengxi YAO ; He YAN ; Wei WEI
International Journal of Traditional Chinese Medicine 2024;46(3):288-292
The clinical manifestations of colorectal polyps are consistent with the characteristics of dampness, stickiness and heaviness. The TCM constitutions in the prone population are mostly related to dampness. The pathological changes of intestinal flora imbalance, intestinal micro inflammation, neuroendocrine immune network and abnormal aquaporin in colorectal polyps are consistent with the research results of modern mechanism of dampness pathogen. This article believed that the TCM pathogenesis of colorectal polyps caused by damp pathogen is the accumulation of spleen deficiency and dampness caused by improper diet, poor emotion and other factors, and the interweaving of various diseases and pathogens to form tangible foreign bodies. According to the pathogenic characteristics of damp pathogen and the pathogenic factors of colorectal polyps, the influence of damp pathogen on the pathogenesis of colorectal polyps was discussed, in order to provide an effective TCM theoretical basis for the diagnosis and treatment of colorectal polyps in clinic.

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