1.Comparative analysis of characteristics and functions of exosomes from human induced pluripotent stem cell-derived platelets and apheresis platelets
Weihua HUANG ; Yan ZANG ; Aihua QIN ; Ziyang FENG ; Heshan TANG ; Fei GUO ; Chuyan WU ; Qiu SHEN ; Baohua QIAN ; Haihui GU ; Zhanshan CHA
Chinese Journal of Blood Transfusion 2025;38(9):1154-1161
Objective: To compare the biological characteristics of human induced pluripotent stem cell-derived platelet exosomes (hiPSC-Plt-Exos) with those of conventional apheresis platelet exosomes (Plt-Exos), specifically focusing on their differential abilities to enhance the proliferation and migration of human umbilical cord mesenchymal stem cells (hUC-MSCs). Methods: Exosomes were isolated from hiPSC-derived Plt and apheresis Plt concentrate using size exclusion chromatography. These exosomes were then characterized through nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and Western blotting. Co-culture experiments into hUC-MSCs were conducted with hiPSC-Plt-Exos and apheresis Plt-Exos, respectively. Their effects on the proliferation and migration of hUC-MSCs were assessed via cell proliferation assays and scratch tests. Results: hiPSC-Plt-Exos and apheresis Plt-Exos exhibited comparable particle sizes, morphological features (such as the characteristic cup-shaped structure), and surface markers (including CD9 and HSP70). Notably, hiPSC-Plt-Exos demonstrated a significantly greater ability to enhance the proliferation and migration of hUC-MSCs compared to apheresis Plt-Exos (P<0.05). These differences provide critical comparative data for their application in various clinical contexts. Conclusion: This study establishes a theoretical foundation for developing precise therapeutic strategies based on hiPSC-Plt-Exos. Furthermore, it underscores the necessity of selecting the appropriate type of exosomes according to the specific disease microenvironment to achieve optimal therapeutic outcomes.
2.Effect of deliberate practice combined with intensive training in cardiopulmonary resuscitation teaching for nurses receiving standardized training
Weiping WANG ; Qiuying DENG ; Chuanren LING ; Weisheng CHEN ; Haihui FANG ; Liqun HUANG
Chinese Journal of Medical Education Research 2024;23(1):123-127
Objective:To investigate the mid- and long-term training effect of deliberate practice combined with intensive training in cardiopulmonary resuscitation (CPR) teaching for nurses receiving standardized training.Methods:A total of 160 nurses who received the first year of standardized training in Guangdong Provincial Hospital of Traditional Chinese Medicine from July 2020 to March 2021 were enrolled as subjects, and they were randomly divided into experimental group and control group, with 80 nurses in each group. The nurses in the control group received the mode of single deliberate practice, and those in the experimental group received deliberate practice combined with intensive training at months 0, 3, and 6. Assessment was performed for the two groups before and after training,with objective indicators and subjective evaluation as the assessment criteria. Objective score was determined by the immediate objective feedback provided by the electronic display of Resusci Anne ? QCPR, and subjective score was determined by the same group of examiners based on the evaluation of cardiopulmonary resuscitation performed by trainees,including environment evaluation, judgment of consciousness, judgment of respiration, initiation of emergency treatment system, re-evaluation after resuscitation, and overall evaluation. SPSS 18.0 was used for the independent samples t-test, the non-parametric test, and the chi-squares test. Results:After training, there were significant differences between the experimental group and the control group in the objective indicators of the percentage of correct compression (CC%) [85.500 (77.50, 93.00) vs. 81.00 (71.75, 89.00), Z=-2.49, P=0.013] and the proportion of correct ventilation (V%) [84.00 (75.00, 92.75) vs. 80.00 (64.00, 90.25), Z=-2.15, P=0.031]. Both groups had significant improvements in objective and subjective indicators after training (all P<0.05). Conclusions:Deliberate practice combined with intensive training at an interval of 3 months can effectively improve the mid- and long-term CPR skills of nurses receiving standardized training and is more suitable for high-level muscle memory training.
3.Rice bran oil supplementation protects swine weanlings against diarrhea and lipopolysaccharide challenge.
Juncheng HUANG ; Wenxia QIN ; Baoyang XU ; Haihui SUN ; Fanghua JING ; Yunzheng XU ; Jianan ZHAO ; Yuwen CHEN ; Libao MA ; Xianghua YAN
Journal of Zhejiang University. Science. B 2023;24(5):430-441
Early weaned piglets suffer from oxidative stress and enteral infection, which usually results in gut microbial dysbiosis, serve diarrhea, and even death. Rice bran oil (RBO), a polyphenol-enriched by-product of rice processing, has been shown to have antioxidant and anti-inflammatory properties both in vivo and in vitro. Here, we ascertained the proper RBO supplementation level, and subsequently determined its effects on lipopolysaccharide (LPS)-induced intestinal dysfunction in weaned piglets. A total of 168 piglets were randomly allocated into four groups of seven replicates (42 piglets each group, (21±1) d of age, body weight (7.60±0.04) kg, and half males and half females) and were given basal diet (Ctrl) or basal diet supplemented with 0.01% (mass fraction) RBO (RBO1), 0.02% RBO (RBO2), or 0.03% RBO (RBO3) for 21 d. Then, seven piglets from the Ctrl and the RBO were treated with LPS (100 μg/kg body weight (BW)) as LPS group and RBO+LPS group, respectively. Meanwhile, seven piglets from the Ctrl were treated with the saline vehicle (Ctrl group). Four hours later, all treated piglets were sacrificed for taking samples of plasma, jejunum tissues, and feces. The results showed that 0.02% was the optimal dose of dietary RBO supplementation based on diarrhea, average daily gain, and average daily feed intake indices in early weaning piglets. Furthermore, RBO protected piglets against LPS-induced jejunal epithelium damage, which was indicated by the increases in villus height, villus height/crypt depth ratio, and Claudin-1 levels, as well as a decreased level of jejunal epithelium apoptosis. RBO also improved the antioxidant ability of LPS-challenged piglets, which was indicated by the elevated concentrations of catalase and superoxide dismutase, and increased total antioxidant capacity, as well as the decreased concentrations of diamine oxidase and malondialdehyde in plasma. Meanwhile, RBO improved the immune function of LPS-challenged weaned piglets, which was indicated by elevated immunoglobulin A (IgA), IgM, β-defensin-1, and lysozyme levels in the plasma. In addition, RBO supplementation improved the LPS challenge-induced dysbiosis of gut microbiota. Particularly, the indices of antioxidant capacity, intestinal damage, and immunity were significantly associated with the RBO-regulated gut microbiota. These findings suggested that 0.02% RBO is a suitable dose to protect against LPS-induced intestinal damage, oxidative stress, and jejunal microbiota dysbiosis in early weaned piglets.
Male
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Female
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Swine
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Animals
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Lipopolysaccharides/toxicity*
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Antioxidants/pharmacology*
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Rice Bran Oil
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Dysbiosis
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Dietary Supplements
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Diarrhea/veterinary*
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Weaning
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Body Weight
4.Efficacy and safety of ixazomib-based therapy for multiple myeloma
Mingxiao YU ; Haihui LIU ; Saisai REN ; Chunyan YANG ; Qian HUANG ; Yanling TAO ; Hao ZHANG
Journal of Leukemia & Lymphoma 2022;31(4):209-212
Objective:To investigate the efficacy and safety of ixazomib-based therapy for multiple myeloma.Methods:The data of 32 patients with multiple myeloma treated with isazomib-based regimen in the Affiliated Hospital of Jining Medical University from December 2020 to December 2021 were retrospectively analyzed. Among 32 patients, 17 cases were relapsed/refractory, and the remaining 15 cases had initial treatment. The treatment regimens included ID (isazomib + dexamethasone), IRD (isazomib + lenalidomide + dexamethasone) and ICD (isazomib + cyclophosphamide + dexamethasone). The short-term curative effect and adverse reactions of relapsed/refractory patients and patients at initial onset were analyzed.Results:The overall response rate (ORR) of relapsed/refractory patients was 52.9% (9/17), of which 6 cases achieved complete remission (CR), 2 cases achieved very good partial remission (VGPR) and 1 case achieved partial remission (PR). The ORR of refractory patients receiving bortezomib therapy was 40.0% (4/10). The ORR of patients at initial onset who could be evaluated the curative effect was 100.0% (14/14), including 9 cases of CR, 2 cases of VGPR and 3 cases of PR. After treatment, 2 patients (6.2%) had grade Ⅲ-Ⅳ adverse events (1 case of herpes zoster and 1 case of thrombocytopenia), and none of the patients had grade Ⅲ-Ⅳ peripheral neuropathy.Conclusion:Isazomib is effective and safe in the treatment of initially treated and relapsed/refractory multiple myeloma.
5.An improved method to generate megakaryocytes from human induced pluripotent stem cells
Weihua HUANG ; Haihui GU ; Yang ZANG ; Yue YANG ; Zhanshan CHA ; Yanxin LI ; Baohua QIAN
Chinese Journal of Blood Transfusion 2022;35(9):900-903
【Objective】 To optimize the existing spin-EB method and promote human induced pluripotent stem cells (hiPSCs) differentiate into megakaryocytes (MKs). 【Methods】 In this study, the initial inoculation amount of hiPSCs was increased from 3 500 cells/well to 8 000 cells/well, and the size of EB was increased. By observing the generation time of EB- hematopoietic cells during differentiation, and detecting the proliferation of CD34+ hematopoietic progenitor cells and CD41+ MKs in different stages, it was studied whether the optimized scheme could promote the differentiation of hiPSCs into hematopoietic progenitor cells(HPCs) and MKs. 【Results】 By increasing the initial inoculation amount of hiPSCs and the size of EB, the differentiation of hiPSCs into HPCs and MKs and the cell production efficiency can be promoted. 【Conclusion】 Our research describes an optimized and repeatable differentiation method, which can produce hematopoietic progenitor cells and mature MKs from hiPSCs in a relatively short time with higher yield. It is of great clinical significance and broad scientific research prospect to continuously optimize the culture scheme of hiPSCs differentiation to produce MKs and platelets in vitro, and to promote large-scale platelet generation in vitro in transfusion medicine.
6.Efficacy analysis of autologous peripheral blood hematopoietic stem cell transplantation in treatment of lymphoma
Jingjing ZHANG ; Yu HUANG ; Qian HUANG ; Lei LIU ; Haihui LIU ; Saisai REN ; Haiyan WANG ; Chunyan YANG ; Lu JIA ; Linlin LYU ; Ying LI ; Dongxiao SONG ; Hao ZHANG
Journal of Leukemia & Lymphoma 2021;30(11):652-657
Objective:To investigate the clinical efficacy of autologous peripheral blood hematopoietic stem cell transplantation (HSCT) in treatment of lymphoma.Methods:The clinical data of 41 lymphoma patients undergoing autologous peripheral blood HSCT at the Affiliated Hospital of Jining Medical University between January 2014 to December 2020 were retrospectively analyzed. There were 6 cases of Hodgkin lymphoma and 35 cases of non-Hodgkin lymphoma. The mobilization regimens included chemotherapy drugs + granulocyte colony-stimulating factor (G-CSF) + thrombopoietin (TPO) or chemotherapy drugs + G-CSF. The pre-conditioning schemes before transplantation were listed as follows: BEAM (mustine + cytarabine + etoposide + melphalan) regimen + decitabine in 26 patients, BEAM regimen in 12 patients, BEAM regimen + chidamide in 3 patients. The progression-free survival (PFS), overall survival (OS), related complications, prognoses after transplantation were observed. The effects of clinical staging, B symptom,International Prognostic Score Index (IPI), extranodal involved sites, hemoglobin (Hb), lactic dehydrogenase (LDH), β 2-microglobulin (β 2-MG), transplantation regimen and the status before transplantation on PFS and OS after transplantation were evaluated. Results:Among 41 patients, 37 patients (90.24%) achieved complete remission (CR), 2 patients (4.88%) achieved partial remission (PR) and 2 patients loss assessment data (4.88%) before autologous peripheral blood HSCT. The median karyocyte count was 12.74×10 8 /kg [(3.91-22.68)×10 8/kg] in 24 patients with the complete data of stem cell collection, the median CD34 positive cell count was 6.74×10 6/kg [(0.91-50.47)×10 6/kg]. All 41 patients had hematologic reconstruction. The median time of platelet implantation was 11 d (7-32 d) and the median time of granulocyte implantation was 9 d (8-16 d). All patients achieved CR after transplantation and no one case had transplantation-related death. By the end of follow-up, 33 cases (80.49%) had no progression of disease, 8 cases (19.51%) died. The OS rates of 12-month, 24-month and 72-month were 93.4%, 85.3% and 60.9%, respectively after transplantation. The PFS rates of 12-month, 24-month and 72 month were 93.3%, 84.0% and 84.0%, respectively. Median PFS and OS had not been reached. There were no statistically significant differences in the PFS and OS of patients with different gender, clinical staging, B symptom, IPI score, extranodal involved sites, Hb, LDH, β 2-MG and the status before transplantation(all P > 0.05) . The PFS and OS of patients receiving BEAM regimen + decitabine were better than those of patients receiving BEAM regimen alone (all P < 0.05). Conclusions:Autologous peripheral blood HSCT is effective in treatment of lymphoma. Moreover, BEAM regimen + dicitabine preconditioning regimen can achieve longer survival time compared with BEAM regimen alone.
7.Research progress on thrombogenesis induced by human stem cells in vitro
Weihua HUANG ; Ruoru WANG ; Haihui GU ; Baohua QIAN
Chinese Journal of Blood Transfusion 2021;34(12):1403-1408
Platelet transfusion is the main clinical treatment of thrombocytopenia. However, due to the difficulty of platelet collection, high cost of use and limited number of blood donors, the development of platelet treatment is greatly limited. Therefore, the research on thrombogenesis in vitro has attracted more attention at home and abroad. Platelet production in vitro has the advantages of donor-independence, platelet antigen free and low risk of alloimmunity. At present, the efficiency of producing functional platelets in vitro is low, and there is still a big gap to achieve the ultimate goal of producing a large number of functional platelets in vitro. This paper reviews the research progress of megakaryocyte / platelet production in vitro, focuses on the in vitro production potential of megakaryocyte / platelet, and summarizes the current platelet culture systems in vitro based on human pluripotent stem cells, embryonic stem cells and adipose stem cells. The contradictions and difficulties of platelet production in vitro were also discussed to provide theoretical support for further research.
8.Interpretation of group standard for Clostridioides difficile infection diagnosis
Yuan WU ; Jinxing LU ; Zhongqiang YAN ; Yunxi LIU ; Wenpeng GU ; Xiaoqing FU ; Yingchun XU ; Anhua WU ; Haihui HUANG ; Zhiyong ZONG ; Dazhi JIN ; Jianhong ZHAO ; Ye CHEN ; Weiping LIU ; Weiguang LI
Chinese Journal of Epidemiology 2021;42(1):64-67
Clostridioides difficile is a key pathogen of antibiotic related diarrhea and hospital associated infection, causing several outbreaks in Europe and North Americans and resulting in severe disease burden. However, the standardized diagnostic principle and detection specifications in C. difficile infection (CDI) survey are limited in China, and the infection rate and disease burden of CDI in China are unclear. Therefore, National Institute for Communicable Disease Control and Prevention,National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, together with another 11 institutions, draft the group standard entitled "Diagnosis of Clostridium difficile infection (T/CPMA 008-2020)" of Chinese Preventive Medicine Association. Based on the principle of "legality, scientificity, advancement, and feasibility", this standard clarifies risk factors, diagnosis principles, diagnoses and differential diagnoses in order to improve the accuracy of CDI diagnosis in clinical practice, guide the surveillance for CDI, and understand the infection rate and disease burden of CDI in China .
9.Relationship between the concentrations of matrix metalloproteinase-9 and adiponectin and postoperative delirium in perioperative serums of elderly patients undergoing lobectomy
Haihui XIE ; Wei DU ; Jianping ZHUO ; Shu ZHANG ; Runcheng HUANG ; Qi HAN
The Journal of Clinical Anesthesiology 2019;35(2):152-155
Objective To evaluate the correlation between the concentrations of matrix metalloproteinase-9 (MMP-9) and adiponectin (ADP) and postoperative delirium (POD) in perioperative serum of elderly patients undergoing lobectomy. Methods Seventy-three elderly patients undergoing lobectomy under general anesthesia, 38 males and 35 females, aged 65-80 years, BMI < 24 kg/m2, falling into ASA physical status Ⅰ or Ⅱ, were selected in some suitable period. All patients were divided into POD group and non-POD group according to the Chinese Version of Consciousness Assessment Scale at 24 hours, 48 hours, and 72 hours after operation, and their blood samples were collected 5 minutes before induction of anesthesia (T0), at the time of tracheal extubation (T1) and at postoperative 24 hours (T2), 48 hours (T3), and 72 hours (T4) to determine the concentrations of MMP-9 and ADP in their serums. Results POD occurred in 19 patients, with the incidence rate of 17.8%. The concentrations of MMP-9 at T1-T4 were significantly higher than those before lobectomy in serums in POD group, while the concentrations of ADP in serums were significantly lower than those before lobectomy (P < 0.05). The concentrations of MMP-9 in serums at T1 were significantly higher than those before lobectomy in non-POD group, while the concentrations of ADP in serums were significantly lower than those before lobectomy (P < 0.05). In the comparison between the two groups, the concentrations of MMP-9 in POD group at T1-T4 were significantly higher than those in non-POD group, while the concentrations of ADP in POD group were significantly lower than those in non-POD group (P < 0.05). Conclusion The serum level of MMP-9 is increased and ADP is decreased in perioperative, wich maybe involved in the pathophysiological process of POD in elderly patients undergoing lobectomy.
10.Applicability of commercial multiplex kit for the diagnosis of: a multicenter investigation Clostridium difficile infections
Shi WU ; Fenfen ZHOU ; Yuxing NI ; Yunjian HU ; Ye CHEN ; Yunsong YU ; Haihui HUANG
Chinese Journal of Laboratory Medicine 2019;42(8):674-678
Objective To evaluate the performance of Xpert C. difficile multiplex real-time PCR assay for diagnosis of Clostridium difficile infections in Chinese hospital settings. Methods This study was performed in Huashan Hospital, Ruijin Hospital, Beijing Hospital, Nanfang Hospital and Sir Run Run Shaw Hospital using a standard study protocol. Unique unformed stools from patients with acute hospital-acquired diarrhea were simultaneously analyzed by toxigenic anaerobic cultures and the Xpert C. difficile assay. All specimens displaying discordant results between the Xpert assay and toxigenic culture were sent for Sanger tcdB gene sequencing. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), total concordance rate, and 95% confidence interval (CI) were calculated before and after resolution of discordant results using SAS 9.3. Results A total of 745 stool specimens were collected and 46 were excluded due to failure of C. difficile recovery. The remaining 699 specimens were included. Compared to the results of toxigenic culture, the sensitivity, specificity, PPV, and NPV of Xpert C. difficile assay were 94.1% (144/153)(95%CI:89.1%-97.3%), 93.2% (509/546)(95%CI:96.7%-99.2%), 79.6% (144/181)(95%CI:72.9%-85.2%)and 98.3% (509 / 518) (95%CI: 96.7%-99.2%), respectively. Both methods had a Kappa of 0.819. Xpert C. difficile assay showed sensitivity of 98.4%(62/63) (95% CI: 90.3%-99.9%) and specificity of 93.2%(509/546) (95% CI: 90.8%-95.2%) for toxin A-negative toxin B-positive strains. After the discordant results resolved by tcdB gene sequencing, PCR assay provided better performance with high sensitivity, specificity, positive predictive value, and negative predictive value [98.8% (171 / 173), 98.1% (516 / 526), 94.5% (171/181) and 99.6% (516/518), respectively]. Conclusions Compared to the results of toxigenic culture, the sensitivity, specificity and NPV of Xpert C. difficile assay were 94.1% (144/153) and 93.2%(509/546), respectively. With the results available within 1 h, Xpert C. difficile assay provides prompt and precise laboratory diagnosis in Chinese clinical settings.

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