1.Design and application effect of continuing education case library combined with case-based learning for rehabilitation therapists
Liguo QIAN ; Tongxuan WU ; Qiaoyun ZHANG ; Jian XING ; Yanyan YANG
Chinese Journal of Rehabilitation Theory and Practice 2026;32(3):249-257
ObjectiveTo investigate the demand and the application outcomes of case-based learning (CBL) combined with teaching case library in continuing education courses for rehabilitation therapists. MethodsA convergent mixed-methods research design was adopted, involving 51 rehabilitation therapists and 31 instructors who participated in the advanced training program at the Department of Rehabilitation Medicine, Peking University Third Hospital between October, 2022 and October, 2024. Self-developed questionnaires were used to collect data on the perceived needs of teachers and students regarding CBL and teaching case library. Differences between CBL + teaching case library and traditional lecturing in student evaluations, classroom participation and interaction were compared using Student Evaluation of Teaching in Medical Lectures, Classroom Participation Scale and Flanders Interaction Analysis System. Semi-structured interviews were conducted to obtain evaluations and attitudes towards this method from both instructors and students' perspectives. ResultsThe survey showed that 91.4% of participating teachers and students supported the use of CBL in the courses, and 82.7% advocated that the teaching case library should include typical cases. Significant differences were observed in teaching preference between teachers and students (χ² = 17.597, P < 0.01). Application effects demonstrated that CBL+teaching library significantly outperformed traditional teaching methods in student previewing behaviors, classroom interaction and learning outcomes (|Z| ≥ 2.646, P < 0.01). Flanders Interaction Analysis indicated that CBL+teaching library was superior to traditional teaching in terms of students' motivation to speak and autonomous learning. Qualitative Research generated four positive themes including cultivating clinical reasoning, being close to clinical practice, deepening knowledge understanding and improving teaching quality; and three negative themes including increasing teaching burden, high software and hardware requirements and posing great challenges to students were generated. ConclusionCompared with traditional teaching methods, CBL combined with teaching case library is closely linked to clinical practice, facilitating students' clinical reasoning, enhancing teaching effectiveness and satisfaction, and therefore aligning with the goals and needs of continuing education for rehabilitation therapists, which is highly recognized by both instructors and students.
2.Design and application effect of continuing education case library combined with case-based learning for rehabilitation therapists
Liguo QIAN ; Tongxuan WU ; Qiaoyun ZHANG ; Jian XING ; Yanyan YANG
Chinese Journal of Rehabilitation Theory and Practice 2026;32(3):249-257
ObjectiveTo investigate the demand and the application outcomes of case-based learning (CBL) combined with teaching case library in continuing education courses for rehabilitation therapists. MethodsA convergent mixed-methods research design was adopted, involving 51 rehabilitation therapists and 31 instructors who participated in the advanced training program at the Department of Rehabilitation Medicine, Peking University Third Hospital between October, 2022 and October, 2024. Self-developed questionnaires were used to collect data on the perceived needs of teachers and students regarding CBL and teaching case library. Differences between CBL + teaching case library and traditional lecturing in student evaluations, classroom participation and interaction were compared using Student Evaluation of Teaching in Medical Lectures, Classroom Participation Scale and Flanders Interaction Analysis System. Semi-structured interviews were conducted to obtain evaluations and attitudes towards this method from both instructors and students' perspectives. ResultsThe survey showed that 91.4% of participating teachers and students supported the use of CBL in the courses, and 82.7% advocated that the teaching case library should include typical cases. Significant differences were observed in teaching preference between teachers and students (χ² = 17.597, P < 0.01). Application effects demonstrated that CBL+teaching library significantly outperformed traditional teaching methods in student previewing behaviors, classroom interaction and learning outcomes (|Z| ≥ 2.646, P < 0.01). Flanders Interaction Analysis indicated that CBL+teaching library was superior to traditional teaching in terms of students' motivation to speak and autonomous learning. Qualitative Research generated four positive themes including cultivating clinical reasoning, being close to clinical practice, deepening knowledge understanding and improving teaching quality; and three negative themes including increasing teaching burden, high software and hardware requirements and posing great challenges to students were generated. ConclusionCompared with traditional teaching methods, CBL combined with teaching case library is closely linked to clinical practice, facilitating students' clinical reasoning, enhancing teaching effectiveness and satisfaction, and therefore aligning with the goals and needs of continuing education for rehabilitation therapists, which is highly recognized by both instructors and students.
3.Design and application effect of continuing education case library combined with case-based learning for rehabilitation therapists
Liguo QIAN ; Tongxuan WU ; Qiaoyun ZHANG ; Jian XING ; Yanyan YANG
Chinese Journal of Rehabilitation Theory and Practice 2026;32(3):249-257
ObjectiveTo investigate the demand and the application outcomes of case-based learning (CBL) combined with teaching case library in continuing education courses for rehabilitation therapists. MethodsA convergent mixed-methods research design was adopted, involving 51 rehabilitation therapists and 31 instructors who participated in the advanced training program at the Department of Rehabilitation Medicine, Peking University Third Hospital between October, 2022 and October, 2024. Self-developed questionnaires were used to collect data on the perceived needs of teachers and students regarding CBL and teaching case library. Differences between CBL + teaching case library and traditional lecturing in student evaluations, classroom participation and interaction were compared using Student Evaluation of Teaching in Medical Lectures, Classroom Participation Scale and Flanders Interaction Analysis System. Semi-structured interviews were conducted to obtain evaluations and attitudes towards this method from both instructors and students' perspectives. ResultsThe survey showed that 91.4% of participating teachers and students supported the use of CBL in the courses, and 82.7% advocated that the teaching case library should include typical cases. Significant differences were observed in teaching preference between teachers and students (χ² = 17.597, P < 0.01). Application effects demonstrated that CBL+teaching library significantly outperformed traditional teaching methods in student previewing behaviors, classroom interaction and learning outcomes (|Z| ≥ 2.646, P < 0.01). Flanders Interaction Analysis indicated that CBL+teaching library was superior to traditional teaching in terms of students' motivation to speak and autonomous learning. Qualitative Research generated four positive themes including cultivating clinical reasoning, being close to clinical practice, deepening knowledge understanding and improving teaching quality; and three negative themes including increasing teaching burden, high software and hardware requirements and posing great challenges to students were generated. ConclusionCompared with traditional teaching methods, CBL combined with teaching case library is closely linked to clinical practice, facilitating students' clinical reasoning, enhancing teaching effectiveness and satisfaction, and therefore aligning with the goals and needs of continuing education for rehabilitation therapists, which is highly recognized by both instructors and students.
4.Exploration on the Effects of Bushen Huoxue Prescription on Rabbit Intervertebral Disc Degeneration Based on the RIPK1/RIPK3/MLKL Signaling Pathway
Wei PENG ; Liguo ZHU ; Xunlu YIN ; Jie LUO ; Kexin YANG ; Minshan FENG ; Jie YU ; Long LIANG ; Linghui LI ; Jiawen ZHAN ; Tao HAN ; Mingyi LUO ; Dian ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(2):71-77
Objective To observe the effects of Bushen Huoxue Prescription on the pathway related to necroptosis of nucleus pulposus cells in a model rabbit of intervertebral disc degeneration;To explore its mechanisms in delaying intervertebral disc degeneration.Methods A intervertebral disc degeneration rabbit model was established using the spinal instability method.Totally 40 model rabbits were randomly divided into model group,ibuprofen group and Bushen Huoxue Prescription low-,medium-and high-dosage groups.Additionally,a normal control group and a sham-operation group were set up,with 8 rabbits in each group.Each treatment groups received the corresponding drugs via gavage for two consecutive weeks.HE staining was used to observe morphology of nucleus pulposus tissue,transmission electron microscopy was used to observe ultrastructure in nucleus pulposus cells,immunohistochemical staining was used to assess the expressions of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue,Western blot was used to detect the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus tissue.Results Compared with the sham-operation group,the model group showed a significant decrease in nucleus pulposus cells,disordered cell arrangement,reduced extracellular matrix,interrupted cell membrane continuity under transmission electron microscopy,organelle swelling,nuclear membrane disruption,partial chromatin loss,and positive expression of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue decreased(P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly increased(P<0.01).Compared with the model group,the treatment groups showed an increased number of nucleus pulposus cells with orderly arrangement and more extracellular matrix,the ultrastructural damage of the cell membrane,organelle and nucleus in nucleus pulposus cells was partially restored under transmission electron microscopy,the positive expressions of Aggrecan and Collagen Ⅱ significantly increased in Bushen Huoxue Prescription medium-and high-dosage groups and the ibuprofen group(P<0.05,P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly decreased(P<0.05,P<0.01).Conclusion Bushen Huoxue Prescription may delay intervertebral disc degeneration of the model rabbit by inhibiting the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus cells,and promoting the generation of extracellular matrix components Aggrecan and Collagen Ⅱ.
5.Exploration on the Effect of Bushen Huoxue Prescription on Necroptosis in Human Nucleus Pulposus Cells Based on RIPK1/RIPK3/MLKL Pathway
Wei PENG ; Liguo ZHU ; Xunlu YIN ; Kexin YANG ; Jie LUO ; Minshan FENG ; Jie YU ; Linghui LI ; Jiawen ZHAN ; Tao HAN ; Long LIANG ; Mingyi LUO ; Dian ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(3):69-75
Objective To observe the effects of Bushen Huoxue Prescription on pressure-induced necroptosis in human nucleus pulposus cells and the expressions of RIPK1/RIPK3/MLKL pathway;To explore its potential mechanism in delaying intervertebral disc degeneration.Methods Human primary nucleus pulposus cells were cultured in vitro,and a model of nucleus pulposus cell degeneration was established using continuous load pressure method.After modeling,the nucleus pulposus cells were divided into model group,Bushen Huoxue Prescription group and inhibitor group,blank serum,Bushen Huoxue Prescription containing serum and necroptotic apoptosis inhibitor(Nec-1)intervention were administered,respectively.Normal group nucleus pulposus cells were cultured routinely.AO/EB fluorescence dual staining method was used for detecting cell apoptosis,flow cytometry was used to detect the necroptosis rate of nucleus pulposus cells,Western blot was used to detect the protein expressions of p-receptor interacting protein kinase(RIPK)1,p-RIPK3 and p-mixed lineage kinase domain like protein(MLKL),RT-qPCR was used to detect the mRNA expressions of RIPK1,RIPK3 and MLKL.Results Compared with the normal group,the model group showed more red fluorescence under AO/EB staining of nucleus pulposus cells,which were round and condensed,the necroptosis rate of nucleus pulposus cells increased(P<0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL increased(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expression increased(P<0.01).Compared with the model group,Bushen Huoxue Prescription group and the inhibitor group had less red condensed chromatin in the nucleus pulposus cells,Bushen Huoxue Prescription group had a lower rate of necroptosis(P<0.05),while the inhibitor group showed a decreasing trend in necroptosis rate(P>0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL decreased in Bushen Huoxue Prescription group and the inhibitor group(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expressions decreased(P<0.01).Conclusion Bushen Huoxue Prescription can alleviate pressure-induced damage to nucleus pulposus cells and inhibit necroptosis,thereby slowing the progression of intervertebral disc degeneration.Its mechanism may be related to the RIPK1/RIPK3/MLKL pathway mediated necroptosis.
6.Analysis of electrocardiogram and echocardiographic characteristics in patients with anti-melanoma differentiation-associated gene 5 antibody-positive dermatomyositis
Ye YUAN ; Zhenwei GUO ; Liguo YIN ; Yanyan BAI ; Jing XU ; Anhao ZHENG ; Shumin ZHANG ; Hongsheng SUN
Chinese Journal of Rheumatology 2025;29(10):855-862
Objective:To study the effect of anti-melanoma differentiation-related gene 5(MDA5) antibody positive dermatomyositis on the heart of patients.Methods:A total of 71 patients with dermatomyositis diagnosed in Shandong Provincial Hospital Affiliated to Shandong First Medical University from January 1, 2014 to December 31, 2019 were enrolled as the sample group, including anti-MDA5 (+) group( n=28); anti-MDA5(-) groups( n=43). Electrocardiogram and echocardiography were performed in the sample group and the control group. The electrocardiogram, echocardiography and other relevant clinical data of the anti-MDA5 (+) group, anti-MDA5 (-) group and the healthy control group were retrospectively analyzed. The logistic regression analysis model was used to analyze the related factors influencing cardiac involvement in anti-MDA5 (+) patients. Results:In the anti-MDA5 (+) group, more than half of the patients showed elevated levels of lactate dehydrogenase (21/28, 75%) and α-hydroxybutyrate dehydrogenase (16/28, 57%), and 11%(3/28) showed elevated levels of creatine kinase isoenzyme and myoglobin. Compared with the anti-MDA5 (-) group, the white blood cell count in the blood routine of the anti-MDA5 (+) group [5.2 (4.0, 6.5) ×10 9/L vs. 7.8 (5.6, 10.6)×10 9/L, Z=-3.447, P=0.001], creatine kinase [62.5 (29.3, 108.3) U/L vs. 481.0 (179.0, 2 738.0) U/L, Z=-5.895, P<0.001], lactate dehydrogenase [313.0 (239.0, 362.0) U/L vs. 448.0 (291.0, 542.0) U/L, Z=-3.236, P<0.001], creatine kinase isoenzyme [1.9 (1.1, 3.9)ng/ml vs. 17.7 (4.0, 67.2) ng/ml, Z=-4.724, P<0.001], myoglobin [28.2 (20.0, 43.0) ng/ml vs. 307.4 (48.1, 612.2) ng/ml, Z=-4.800, P<0.001]. Electrocardiogram analysis showed that QRS axis [33.5±265.9 vs. 46.9±22.4, t=-2.900, P=0.004], SV1 amplitude [0.7 (0.4, 0.9) vs. 0.9 (0.7, 1.0), Z=-2.148, P=0.023] in anti-MDA5 antibody (+) group in anti-MDA5 antibody (+) group were lower than anti-MDA5 antibody (-) group. QRS duration [84.0 (78.0, 96.5) vs.92.0 (87.8, 100.5), Z=-2.900, P=0.004], QRS axis [33.5±265.9 vs. 46.9±20.4, Z=-2.32, P=0.023] in the anti-MDA5 antibody (+) group were lower than those in healthy control group. Echocardiographic analysis showed that the E peak of anti-MDA5 (+) group [63.0 (52.5, 69.5)] was significantly lower than that of anti-MDA5 (-) group [85.0 (68.0, 108.0), Z=-4.926, P<0.001)]and healthy control group [67.0 (62.8, 80.3), Z=-2.897, P=0.004]. The peak A of anti-MDA5 (+) group [65.5 (56.5, 80.0)] was significantly lower than that of anti-MDA5 (-) group [76.0 (65.0, 90.0), Z=-2.631, P=0.011], but higher than that of healthy control group [55.0(51.0, 66.5), Z=-4.550, P<0.001]. There was no significant difference in echocardiographic findi-ngs among the other groups. All patients with anti-MDA5 (+) dermatomyositis had interstitial lung disease (28/28, 100%). Patients with MDA5 antibody (+++) are more likely to have cardiac involvement than patients with MDA5 antibody (++). Conclusion:The results of relevant examinations in anti-MDA5-DM patients suggest that there is damage to myocardial cells and cardiac function.
7.Short-term outcome study on cervical deep lymph node-venous anastomosis technique in the treatment of Alzheimer’s disease
Cheng GAN ; Zhengdong KONG ; Xiaoye RAN ; Shudong QIAO ; Yixin ZHANG ; Lu YUE ; Yingjie WANG ; Hui BI ; Dong YANG ; Hongtong MA ; Yuan CHEN ; Hongli CHAI ; Ying JIA ; Chenhao MA ; Zixiang CHEN ; Ke LI ; Miao WANG ; Liguo XUE ; Siwen ZHAO ; Ke WEN ; Lin YIN ; Bo DING ; Shan ZHU ; Yuanbo LIU ; Mengqing ZANG
Chinese Journal of Plastic Surgery 2025;41(2):130-143
Objective:To explore the short-term clinical effects of deep cervical lymph node-venous anastomosis in the treatment of Alzheimer’s disease (AD).Methods:A prospective exploratory study was conducted on the treatment of AD patients using the cervical deep lymph node-venous anastomosis technique in Scar and Wound Treatment Department, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, from September to October 2024. The patients underwent high-frequency ultrasound to locate deep cervical lymph nodes and the external jugular vein. Under general anesthesia, bilateral deep cervical lymph node-venous anastomoses were performed. Indocyanine green (ICG) lymphography was conducted via subcutaneous injection behind the ear to visualize lymph nodes in levels Ⅱ and Ⅲ. After making a skin incision along the posterior margin of the sternocleidomastoid muscle, the external jugular vein, internal jugular veins, and associated lymph nodes were exposed. Adjacent veins were selected for anastomosis of lymph node. Using microsurgical techniques, end-to-side or end-to-end anastomosis was completed for lymph nodes in levels Ⅱ and Ⅲ. Preoperative assessments included the mini-mental state examination (MMSE, a higher score indicates better cognitive function), Alzheimer’s disease assessment scale-cognitive subscale (ADAS-Cog, a higher score indicates greater impairment of cognitive function), Alzheimer’s disease cooperative study scale for activities of daily living (ADCS-ADL, a higher score indicates better ability to perform daily activity), and neuropsychiatric inventory (NPI, a higher score indicates more severe behavioral and emotional symptom). Postoperative follow-up included the same scales to observe changes in cognitive function, activities of daily living, and emotional communication.Results:Four patients (1 male, 3 females, aged 58-79 years) with AD were included. All were diagnosed based on cerebrospinal fluid biomarkers. All patients successfully underwent bilateral deep cervical lymph node-venous anastomoses. On average, 4.3 (2-7 per person) anastomoses were performed per patient. Surgical procedures lasted an average of 6.5 h (5.5-8.5 h) with minimal blood loss (less than 50 ml). Patients resumed normal activity within 6 hours postoperatively and were discharged after an average of 4.1 d (3.5-5.0 d). Postoperative complications included one case each of aspiration pneumonia, lower limb venous thrombosis, and transient delirium, all of whom resolved without long-term effects. Clinical symptoms, including memory decline, mood swings, and anxiety, showed varying degrees of improvement. Patients reported enhanced quality of life, emotional stability, and social engagement, confirming the procedure’s safety and potential cognitive benefits. At one month postoperatively, the MMSE scores of the four patients increased by an average of 0.8 points compared to preoperative levels. Additionally, the two patients who completed the ADAS-Cog assessments showed a decrease in their scores (reduced by 1.0 points and 11.3 points, respectively, compared to preoperative scores), indicating a certain degree of improvement in cognitive function during this period. The ADCS-ADL and NPI scores of four patients varied significantly, without showing any clear pattern.Conclusion:Lymphovenous anastomosis of the deep cervical lymph node-venous anastomosis may provide a new surgical intervention approach for AD, but further large-scale studies and long-term follow-up are needed to validate its safety and effectiveness.
8.Mechanotransduction proteins in intervertebral disc degeneration
Xilin GAO ; Si WU ; Chao ZHANG ; Liguo ZHU ; Bifeng FU ; Ping WANG
Chinese Journal of Tissue Engineering Research 2025;29(3):579-589
BACKGROUND:Recent research indicates that disc degeneration is closely related to abnormal stress load,and mechanotransduction proteins play a key role in it. OBJECTIVE:To investigate the role and mechanism of mechanotransduction proteins in the mechanotransduction process induced by abnormal mechanical stimulation in disc degeneration,and to summarize the current treatment strategies targeting mechanotransduction to delay intervertebral disc degeneration. METHODS:Using"intervertebral disc,nucleus pulposus,annulus fibrosus,cartilaginous endplate,cell,mechanics,signal transduction,protein,biomechanics"as Chinese search terms,and"intervertebral disc,nucleus pulposus,annulus fibrosus,cartilaginous endplate,cell,mechanical stimulation,signal transduction,protein,biomechanics"as English search terms,relevant literature in the PubMed and CNKI databases was searched.A total of 88 articles were ultimately included for review. RESULTS AND CONCLUSION:Disc cells can sense external mechanical stimulation through various mechanotransduction proteins and convert it into biological responses within the cells.These transduction proteins mainly include collagen proteins in the extracellular matrix,cell membrane surface receptors(such as integrins and ion channels),and cytoskeleton structural proteins.Their regulation of mechanotransduction processes primarily involves the activation of multiple pathways,such as the PI3K/AKT signaling pathway,nuclear factor-kB signaling pathway,and Ca2+/Calpain2/Caspase3 pathway.Mechanotransduction proteins play a key role in the mechanotransduction of disc cells.Abnormal expression of these proteins or resulting changes in the extracellular matrix environment can disrupt the mechanical balance of disc cells,leading to disc degeneration.In-depth study of the expression and regulatory mechanisms of mechanotransduction proteins in disc cells,and identification of key pathological links and therapeutic targets,is of significant importance for developing treatment strategies for disc degeneration.Current strategies to delay intervertebral disc degeneration by targeting mechanotransduction mainly include regulation of transduction proteins and improvement of the extracellular matrix.However,research in this area is still in its early stages.As research continues,new breakthroughs are expected in the regulation of disc degeneration by mechanotransduction proteins.
9.Expert consensus on the positioning of the "Three-in-One" Registration and Evaluation Evidence System and the value of orientation of the "personal experience"
Qi WANG ; Yongyan WANG ; Wei XIAO ; Jinzhou TIAN ; Shilin CHEN ; Liguo ZHU ; Guangrong SUN ; Daning ZHANG ; Daihan ZHOU ; Guoqiang MEI ; Baofan SHEN ; Qingguo WANG ; Xixing WANG ; Zheng NAN ; Mingxiang HAN ; Yue GAO ; Xiaohe XIAO ; Xiaobo SUN ; Kaiwen HU ; Liqun JIA ; Li FENG ; Chengyu WU ; Xia DING
Journal of Beijing University of Traditional Chinese Medicine 2025;48(4):445-450
Traditional Chinese Medicine (TCM), as a treasure of the Chinese nation, plays a significant role in maintaining public health. In 2019, the Central Committee of the Communist Party of China and the State Council proposed for the first time the establishment of a TCM registration and evaluation evidence system that integrates TCM theory, "personal experience" and clinical trials (referred to as the "Three-in-One" System) to promote the inheritance and innovation of TCM. Subsequently, the National Medical Products Administration issued several guiding principles to advance the improvement and implementation of this system. Owing to the complexity of its implementation, there are still differing understandings within the TCM industry regarding the positioning of the "Three-in-One" Registration and Evaluation Evidence System, as well as the connotation and value orientation of the "personal experience." To address this, Academician WANG Qi, President of the TCM Association, China International Exchange and Promotion Association for Medical and Healthcare and TCM master, led a group of academicians, TCM masters, TCM pharmacology experts and clinical TCM experts to convene a "Seminar on Promoting the Implementation of the ′Three-in-One′ Registration and Evaluation Evidence System for Chinese Medicinals." Through extensive discussions, an expert consensus was formed, clarifying the different roles of the TCM theory, "personal experience" and clinical trials within the system. It was further emphasized that the "personal experience" is the core of this system, and its data should be derived from clinical practice scenarios. In the future, the improvement of this system will require collaborative efforts across multiple fields to promote the high-quality development of the Chinese medicinal industry.
10.High-throughput single-microbe RNA sequencing reveals adaptive state heterogeneity and host-phage activity associations in human gut microbiome.
Yifei SHEN ; Qinghong QIAN ; Liguo DING ; Wenxin QU ; Tianyu ZHANG ; Mengdi SONG ; Yingjuan HUANG ; Mengting WANG ; Ziye XU ; Jiaye CHEN ; Ling DONG ; Hongyu CHEN ; Enhui SHEN ; Shufa ZHENG ; Yu CHEN ; Jiong LIU ; Longjiang FAN ; Yongcheng WANG
Protein & Cell 2025;16(3):211-226
Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications for health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve a comprehensive understanding of complex microbial communities together with their hosts are therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive response states among species in Prevotella and Roseburia genera and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated that smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-world situations and promises new perspectives in the understanding of human microbiomes.
Humans
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Gastrointestinal Microbiome/genetics*
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Bacteriophages/physiology*
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High-Throughput Nucleotide Sequencing
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Sequence Analysis, RNA/methods*
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Bacteria/virology*


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