1.Research on Development Path and Strategy of Human Use Experience in Traditional Chinese Medicine Based on Bibliometrics and Thematic Analysis
Yundan WU ; Qun CHEN ; Jie CHEN ; Yuhang OU ; Jindong WU ; Yan XIAO ; Jiemei GUO ; Jing CAI ; Youxin SU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):118-128
ObjectiveThe development trend and knowledge structure of the research on human use experience (HUE) of traditional Chinese medicine (TCM) were systematically reviewed, and the core challenges and future directions were identified. This study aims to provide reference for the construction of a scientific and feasible research and development framework and evidence transformation system. MethodsLiterature related to "human use experience" published from January 1, 2019 to July 31, 2025 was retrieved from the China National Knowledge Infrastructure (CNKI), Wanfang, China Science and Technology Journal Database (VIP), and PubMed databases. Bibliometric visualization was conducted using Excel, VOSviewer, and CiteSpace, followed by in-depth reading and thematic summarization of core literature. ResultsA total of 181 papers were included for bibliometric analysis, with 45 articles used for in-depth thematic mining. The analysis showed that the number of publications on HUE research has increased in a stepwise manner over the past five years. Yang Zhongqi (24 times) was the core of the author network, the journal with the highest number of publications was China Journal of Chinese Materia Medica, the institutions publishing the most articles were mainly research institutions, regulatory agencies, hospitals, and universities, high-frequency keywords included "new TCM drugs", "real-world studies", and "clinical comprehensive evaluation", keyword clustering analysis formed three major clusters: Policy orientation, application fields, and methodological approaches. Thematic analysis reveals that HUE-based evaluation should be integrated throughout the research and development process, encompassing three dimensions: TCM theory, clinical value, and pharmaceutical fundamentals, with toxic herbs and compatibility contraindications being key foci. Data collection primarily relies on empirical data, while real-world data constitute the primary source for clinical research, with efficacy and safety as the shared core. Data management emphasizes quality control and statistical analysis; however, the management of bias and confounding remains a critical bottleneck in evidence transformation. In practice, HUE-based approaches have successfully supported the registration and evaluation of multiple categories of new TCM drugs. ConclusionThe research on HUE of TCM has formed a policy-driven pattern characterized by, rapid development and close link with regulatory practice. A technical framework covering the whole chain of research and development has been constructed with clinical value as the core, which provides methodological basis and strategy reference for the scientific transformation of HUE of TCM from "experience" to "evidence".
2.Impact of birth weight on the trajectory of blood pressure among primary school students
CUI Chengpeng, YE Siyan, FANG Yanfei, LI Yan, PENG Zeqin, XIAO Yuqing, WU Meng, LIU Qin
Chinese Journal of School Health 2026;47(3):309-313
Objective:
To explore the early effects of birth weight at different gestational ages on blood pressure trajectory among primary school students, so as to provide evidence for incorporating gestational age birth weight into individualized early warning and intervention strategies for childhood hypertension.
Methods:
From May to November 2023, a purposeful sampling method was used to recruit 1 676 students in grade 1-3 from three primary schools in a certain urban district of Chongqing. Follow up assessments were conducted in May 2024(T1), November 2024(T2), and May 2025(T3). General demographic and birth related information were collected via self administered questionnaires, while height, weight and blood pressure were obtained through physical examinations. Linear mixed effects model was used to analyze the associations between birth weight at different gestational ages and blood pressure trajectories.
Results:
During the T1-T3 period, the systolic blood pressure of boys were 98.5 (93.0, 104.5 ),98.5 (93.5, 105.0), and 97.5 (92.5, 103.5)mmHg, respectively, while the diastolic blood pressure were 60.5 (56.5, 65.0), 61.5 ( 57.0 , 65.5), and 60.0 (56.0, 64.0)mmHg, respectively. For girls, the systolic blood pressure were 95.5 (90.0, 102.0),95.5 (90.5, 101.5), and 96.0 (90.5, 101.5)mmHg, respectively, and the diastolic blood pressure were 60.5 (56.0, 64.5 ),61.5 (57.5, 65.5), and 59.5 (56.0, 63.0)mmHg, respectively. Through Friedman test within both boys and girls, diostolic blood pressure were statistically significant across three measurements( χ 2=48.85,81.54,both P <0.01). The proportion of normal blood pressure increased , and the proportion of prehypertension and hypertension decreased with time( χ 2=39.72,25.62,both P < 0.01 ). Linear mixed effects model analysis revealed that after adjusting for age, sex, household income monthly, parental education, family history of hypertension and maternal pregnancy complications, large for gestational age had significantly higher trajectories of systolic ( β = 1.50) and diastolic( β =0.94) blood pressure compared to appropriate for gestational age(both P <0.01).
Conclusion
Large for gestational age is associated with elevated blood pressure trajectories during school age, and it may be considered as an early indicator for individualized screening and intervention for childhood hypertension.
3.Clinical Advantages of Traditional Chinese Medicine in Treatment of Childhood Simple Obesity: Insights from Expert Consensus
Qi ZHANG ; Yingke LIU ; Xiaoxiao ZHANG ; Guichen NI ; Heyin XIAO ; Junhong WANG ; Liqun WU ; Zhanfeng YAN ; Kundi WANG ; Jiajia CHEN ; Hong ZHENG ; Xinying GAO ; Liya WEI ; Qiang HE ; Qian ZHAO ; Huimin SU ; Zhaolan LIU ; Dafeng LONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):238-245
Childhood simple obesity has become a significant public health issue in China. Modern medicine primarily relies on lifestyle interventions and often suffers from poor long-term compliance, while pharmacological options are limited and associated with potential adverse effects. Traditional Chinese Medicine (TCM) has a long history in the prevention and management of this condition, demonstrating eight distinct advantages, including systematic theoretical foundation, diversified therapeutic approaches, definite therapeutic efficacy, high safety profile, good patient compliance, comprehensive intervention strategies, emphasis on prevention, and stepwise treatment protocols. Additionally, TCM is characterized by six distinctive features: the use of natural medicinal substances, non-invasive external therapies, integration of medicinal dietetics, simple exercise regimens, precise syndrome differentiation, and diverse dosage forms. By combining internal and external treatments, TCM facilitates individualized regimen adjustment and holistic regulation, demonstrating remarkable effects in improving obesity-related metabolic indicators, regulating constitutional imbalance, and promoting healthy behaviors. However, challenges remain, such as inconsistent operational standards, insufficient high-quality clinical evidence, and a gap between basic research and clinical application. Future efforts should focus on accelerating the standardization of TCM diagnosis and treatment, conducting multicenter randomized controlled trials, and fostering interdisciplinary integration, so as to enhance the scientific validity and international recognition of TCM in the prevention and treatment of childhood obesity.
4.Shaoyaotang Regulates miRNA-155-mediated SOCS1/JAK1/STAT1 Signaling Pathway to Affect Macrophage Polarization
Qi CHENG ; Bo ZOU ; Youwei XIAO ; Yiqian YU ; Ruoru HUANG ; Yan GONG ; Jiachun XIONG ; Jun XIONG ; Dichang LAI ; Dongsheng WU ; Hui CAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):43-52
ObjectiveTo investigate the mechanism by which Shaoyaotang regulates the miRNA-155-mediated suppressor of cytokine signaling 1 (SOCS1)/Janus kinase 1 (JAK1)/signal transducer and activator of transcription 1 (STAT1) signaling pathway and thereby affects macrophage polarization. MethodsThe cell-counting kit-8 (CCK-8) assay was used to detect the effect of drug-containing serum of Shaoyaotang at different concentrations on the viability of RAW 264.7 cells. A cell model of inflammation was established by stimulating RAW264.7 cells with lipopolysaccharide (LPS) at a concentration of 10 mg·L-1 The modeled cells were assigned by the random number table method into seven groups: LPS-induced M1 polarization (model), M1+miRNA-155 mimics, M1+miRNA-155 inhibitor, M1+Shaoyaotang-containing serum, M1+miRNA-155 mimics+Shaoyaotang-containing serum, M1+miRNA-155 inhibitor+Shaoyaotang-containing serum, and M1+blank serum. Enzyme-linked immunosorbent assay was employed to measure the levels of inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β)]. Immunofluorescence assay was used to detect the expression of macrophage polarization markers [inducible nitric oxide synthase (iNOS) and macrophage mannose receptor 1 (CD206)]. Real-time PCR was employed to measure the expression of miRNA-155 in cells. Western blot was performed to determine the protein levels of SOCS1, STAT1, and JAK1. ResultsCompared with the LPS-induced M1 polarization (model) group, the M1+miRNA-155 mimics group showed up-regulated expression of miRNA-155, JAK1, STAT1, TNF-α, IL-6, IL-1β, and iNOS (P<0.05) and down-regulated expression of CD206 (P<0.05). In both the M1+miRNA-155 inhibitor group and the M1+Shaoyaotang-containing serum group, the expression levels of miRNA-155, JAK1, STAT1, TNF-α, IL-6, IL-1β, and iNOS were down-regulated (P<0.05), while those of SOCS1 and CD206 were up-regulated (P<0.05). Compared with the M1+miRNA-155 mimics group, the M1+miRNA-155 mimics+Shaoyaotang-containing serum group showed down-regulated expression of miRNA-155, JAK1, STAT1, TNF-α, IL-6, IL-1β, and iNOS (P<0.05) and up-regulated expression of SOCS1 and CD206 (P<0.05). Compared with the M1+miRNA-155 inhibitor group, the M1+miRNA-155 inhibitor+Shaoyaotang-containing serum group showed down-regulated expression of miRNA-155, JAK1, STAT1, TNF-α, IL-6, IL-1β, and iNOS (P<0.05) and up-regulated expression of SOCS1 and CD206 (P<0.05). ConclusionShaoyaotang regulates macrophage polarization by modulating miRNA-155 expression and interfering with the SOCS1/JAK1/STAT1 signaling pathway. The findings provide new experimental evidence for the treatment of ulcerative colitis with Shaoyaotang.
5.Effect and Mechanisms of Shaoyaotang on Murine Ulcerative Colitis via Modulating Macrophage Glycolytic Reprogramming and Polarization Through HIF-1α Pathway
Yiqian YU ; Hui CAO ; Dongsheng WU ; Bo ZOU ; Ruoru HUANG ; Qi CHENG ; Youwei XIAO ; Yan GONG ; Jiachun XIONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):53-60
ObjectiveTo investigate the potential role and underlying mechanisms of Shaoyaotang in intervening macrophage glycolytic reprogramming in ulcerative colitis (UC). MethodsForty-eight C57BL/6 mice were randomly divided into six groups: Normal control group, model group, mesalazine group (0.39 g·kg-1), Shaoyaotang group (15.54 g·kg-1), 2-deoxy-D-glucose (2-DG) group (glycolysis inhibitor, 100 mg·kg-1), and 2-DG + Shaoyaotang combined group (100 mg·kg-1+15.54 g·kg-1). Except for the normal control group, mice in the other five groups were induced to establish UC models using dextran sulfate sodium (DSS). The normal control group was administered pure water via intragastric gavage, while the other groups received intragastric gavage of mesalazine solution, intragastric gavage of Shaoyaotang, and the 2-DG group was treated with 2-DG via intraperitoneal injection. After 7 consecutive days of treatment, colonic tissues were extracted. Hematoxylin and eosin (HE) staining was performed to evaluate histopathological changes and tissue injury in the colon. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in colonic tissues. Western blot analysis was employed to determine the expression levels of hypoxia-inducible factor-1α (HIF-1α), glucose transporter (GLUT1), lactate dehydrogenase A (LDHA), pyruvate kinase M2 (PKM2), and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) in colonic tissues. Immunofluorescence was conducted to detect the expression of CD206 and inducible nitric oxide synthase (iNOS) in colonic tissues. Liquid chromatography-mass spectrometry (LC-MS) was utilized to measure lactate and citrate levels in colonic tissues. ResultsCompared with the normal control group, mice in the model group exhibited a significant increase in disease activity index (DAI) scores, accompanied by colonic mucosal congestion, edema, and inflammatory cell infiltration, significantly elevated expression of the inflammatory cytokine TNF-α (P<0.05), significantly decreased IL-10 expression (P<0.05), significantly increased levels of HIF-1α, GLUT1, LDHA, PKM2, and PFKFB3 in colonic tissues (P<0.05), markedly elevated iNOS expression (P<0.05), significantly decreased CD206 expression (P<0.05), and significantly elevated lactate and citrate levels in colonic tissues (P<0.05). In contrast to the model group, the Shaoyaotang group, inhibitor group, and Shaoyaotang combined with inhibitor group demonstrated amelioration of mucosal injury in colonic tissues, markely decreased expression levels of the inflammatory cytokine TNF-α (P<0.05), elevated IL-10 expression levels, significantly decreased expression of HIF-1α, GLUT1, LDHA, PKM2, and PFKFB3 (P<0.05), markedly reduced iNOS expression levels (P<0.05), significantly increased CD206 expression (P<0.05) and significantly decreased lactate and citrate levels (P<0.05). ConclusionShaoyaotang ameliorates symptoms of DSS-induced UC in mice, and its therapeutic mechanism may be associated with regulating macrophage glycolytic reprogramming via modulation of the HIF-1α signaling pathway.
6.Shaoyaotang Ameliorates Ulcerative Colitis by Regulating miR-155-5p
Ruoru HUANG ; Bo ZOU ; Yu ZHANG ; Yiqian YU ; Qi CHENG ; Youwei XIAO ; Jiachun XIONG ; Yan GONG ; Dongshen WU ; Hui CAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):61-68
ObjectiveTo investigate the role of microRNA-155-5p (miR-155-5p) in ulcerative colitis (UC) and study the molecular mechanism of Shaoyaotang in the treatment of UC by regulating miR-155-5p. MethodsForty-eight SPF-grade male C57BL/6 mice were selected and assigned via the random number table method into 6 groups (n=8): A blank control group, a model group, a mesalazine (0.39 g·kg-1) group, a Shaoyaotang (31.08 g·kg-1) group, a Janus kinase 1 (JAK1) inhibitor (baricitinib, 10 mg·kg-1) group, and a Shaoyaotang combined with inhibitor (baricitinib 10 mg·kg-1 + Shaoyaotang 31.08 g·kg-1) group. After successful modeling of UC by gavage of 3% dextran sulphate sodium solution, each group received corresponding drug intervention for 7 days. Shaoyaotang and mesalazine were administered by gavage, and baricitinib by intraperitoneal injection. Twenty-four hours after the last administration, mice were anesthetized by intraperitoneal injection of pentobarbital sodium, and blood was collected for determination of white blood cell count and erythrocyte sedimentation rate (ESR). Mice were then sacrificed for measurement of colon length. Hematoxylin-eosin staining was used to observe colonic pathological changes and perform pathological scoring. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was employed to determine the relative expression of miR-155-5p in the colonic tissue, and Western blot was used to determine the protein levels of JAK1, phosphorylated JAK1 (p-JAK1), suppressor of cytokine signaling 1 (SOCS1), signal transducer and activator of transcription 1 (STAT1), and phosphorylated STAT1 (p-STAT1). ResultsCompared with the blank control group, the model group showed increased disease activity index (DAI) score and pathological score, shortened colon, upregulated relative expression of miR-155-5p and protein levels of p-JAK1 and p-STAT1, downregulated protein level of SOCS1 in the colonic tissue, prolonged time of erythrocyte sedimentation, and increased white blood cell count (P<0.01). Compared with the model group, all drug-treated groups exhibited improvements in the above indicators (P<0.01). Moreover, the Shaoyaotang group showed better therapeutic effects than the mesalazine group in regulating miR-155-5p expression, related protein levels, DAI score, and colonic pathological score (P<0.01). ConclusionShaoyaotang may downregulate miR-155-5p to relieve its inhibition on SOCS1, thereby suppressing the excessive activation of the JAK1/STAT1 signaling pathway and ultimately alleviating intestinal inflammatory damage.
7.Shaoyaotang Regulates TLR4/MyD88/NF-κB Signaling Pathway to Protect Intestinal Mucosal Barrier in Ulcerative Colitis
Dongsheng WU ; Yu ZHANG ; Wenjing QUAN ; Wanqing XIONG ; Bo ZOU ; Youwei XIAO ; Ruoru HUANG ; Yan GONG ; Hui CAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):69-75
ObjectiveTo investigate the role of the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway in intestinal mucosal barrier damage in ulcerative colitis, as well as the intervention mechanism of Shaoyaotang. MethodsSixty SD rats were allocated into a blank group, a model group, a mesalazine (0.42 g·kg-1) group, and low-, medium-, and high-dose (11.1, 22.2, 44.4 g·kg-1, respectively) Shaoyaotang groups. A model of ulcerative colitis was induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS). After successful modeling, rats were administrated with corresponding agents via gavage for 7 days. Changes in colon length and colon weight were observed. Hematoxylin-eosin staining was performed to examine the pathological changes of the colon, and immunohistochemistry was employed to detect the expression of the inflammatory cytokine interleukin-8 (IL-8), cyclooxygenase-2 (COX-2), junction adhesion molecule-1 (JAM-1), and claudin-1 in the colon. Western blot analysis was performed to determine the protein levels of TLR4, MyD88, and NF-κB in the colon. ResultsCompared with the blank group, the model group showed elevated DAI score (P<0.01), reduced colon length and colon weight (P<0.01), down-regulated protein levels of JAM-1 and claudin-1 (P<0.01), and up-regulated protein levels of IL-8, COX-2, TLR4, MyD88, and NF-κB p65 (P<0.01) in the colon tissue. Compared with the model group, each treatment group showed decreased DAI score (P<0.05, P<0.01), increased colon length and colon weight (P<0.05, P<0.01), up-regulated protein levels of JAM-1 and claudin-1 (P<0.01), and down-regulated protein levels of IL-8, COX-2, TLR4, MyD88, and NF-κB p65 (P<0.01) in the colon tissue. ConclusionShaoyaotang alleviates intestinal inflammation and intestinal mucosal damage to protect intestinal barrier integrity by regulating the TLR4/MyD88/NF-κB signaling pathway.
8.Salidroside alleviates progression of Parkinson's disease by modulating inflammatory responses
Xiao-lin DONG ; Gang WU ; Yan-ping LI ; Li-juan ZHANG ; Fu-rong JIN ; Rui LI ; Hong-mei LI ; Xiao-xiao ZHANG ; Qing-yun LI
Chinese Pharmacological Bulletin 2025;41(7):1340-1345
Aim To explore the neuroprotective effects of salidroside on Parkinson's disease(PD)through modulation of inflammatory responses and the underly-ing mechanisms.Methods Mice were divided into five groups:healthy control group,1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)disease group,low-dose Rhodioloside intervention group,medium-dose salidroside intervention group,and high-dose salidro-side intervention group.MPTP-induced PD mouse model was established,and salidroside intervention was administered.Behavioral changes,inflammatory cyto-kine levels,autophagy-related protein expression,and neurons were observed through histological analysis and immunohistochemical staining.Results After MPTP treatment,mice exhibited significant behavioral chan-ges,increased pro-inflammatory cytokines,decreased anti-inflammatory cytokines,reduced autophagy-related proteins,and evident pyroptosis.Salidroside interven-tion alleviated these changes in a dose-dependent man-ner.Conclusions Salidroside exerts neuroprotective effects on PD by alleviating inflammatory responses and promoting autophagy,thereby protecting neurons.
9.Analysis of completion rate of tumor evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer : a national multicenter real-world study
Kexuan LI ; Tixian XIAO ; Xiaodong WANG ; Bin WU ; Guole LIN ; Yuchen GUO ; Ming QU ; Si WU ; Xiaodong YANG ; Yinshengbo′er BAO ; Baohua WANG ; Fan ZHANG ; Xiangwang YU ; Beizhan NIU ; Junyang LU ; Lai XU ; Guannan ZHANG ; Zhen SUN ; Guoyou ZHANG ; Yan SHI ; Hong JIANG ; Yongjing TIAN ; Yongxiang LI ; Hongwei YAO ; Jun XUE ; Quan WANG ; Lie YANG ; Qian LIU ; Yi XIAO
Chinese Journal of Digestive Surgery 2025;24(1):113-119
Objective:To investigate the completion rate of tumor evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer patients in the national multicenter real-world database.Methods:The prospective real-world study was conducted. The clinicopathological data of 1 074 patients who underwent surgical treatment for mid and low rectal cancer in 47 national medical institutions, including Peking Union Medical College Hospital et al, from May 12,2023 to May 11,2024 were collected. Observation indicators: (1) clinical characteristics of patients with mid and low rectal cancer; (2) initial colonoscopy and pathologic evaluation of tumors in patients with mid and low rectal cancer; (3) initial imaging evaluation of patients with mid and low rectal cancer; (4) imaging evaluation after neoadjuvant therapy for patients with mid and low rectal cancer. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M( Q1, Q3). Count data were described as absoluter numbers and/or percentages. Results:(1) Clinical characteristics of patients with mid and low rectal cancer. Of the 1 074 patients, there were 713 males and 361 females, aged 63(56,70)years. The body mass index of 1 074 patients was 24(21,26)kg/m 2.For American Society of Anesthesiologists classification, there were 147 cases of stage Ⅰ, 641 cases of stage Ⅱ, 157 cases of stage Ⅲ, 2 cases of stage Ⅳ, and there were 127 cases missing data. (2) Initial colonoscopy and pathologic evaluation of tumors in patients with mid and low rectal cancer. Of the 1 074 patients, there were 787 cases (73.28%) undergoing complete colonoscopy, and there were only 197 cases (18.34%) undergoing immunohistochemical evaluation of all four mismatch repair proteins. (3) Initial imaging evaluation of patients with mid and low rectal cancer. Of the 1 074 patients, there were 842(78.40%) patients completing magnetic resonance imaging (MRI) or ultrasound evaluation, and there were 914(85.10%) patients completing chest, abdomen, and pelvis enhanced computed tomography (CT) evaluation. In the 149 patients completing rectal ultrasound evaluation, there were 122 cases (81.88%) comple-ting T staging evaluation, and there were 81 cases (54.36%) completing N staging evaluation. In the 808 patients completing rectal MRI evaluation, there were 708 cases (87.62%) completing T staging evaluation, and there were 590 cases (73.02%) completing N staging evaluation. (4) Imaging evalua-tion after neoadjuvant therapy for patients with mid and low rectal cancer. Of the 388 patients with neoadjuvant therapy, there were 332 patients (85.57%) completing MRI or ultrasound evaluation, and there were 327 patients (84.28%) completing chest, abdomen, and pelvis enhanced CT evalua-tion. In the 70 patients completing rectal ultrasound evaluation, there were 65 cases (92.86%) com-pleting T staging evaluation, and there were 49 cases (70.00%) completing N staging evaluation. In the 327 patients completing rectal MRI evaluation, there were 246 cases (75.23%) completing T staging, and there were 228 cases (69.72%) completing N staging evaluation. Conclusion:The com-pletion rate of tumor imaging evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer patients on a national scale is relatively good.
10.Atomic force microscopic observation of surface structure of purified protein molecules in vitro
Zhao-ying CHEN ; Xiao-feng FANG ; Dan-yan LIU ; Meng-qin CHEN ; Xi LING ; Jie-min ZHAO ; Wei WEI ; Yu-jing WU
Chinese Pharmacological Bulletin 2025;41(1):193-199
Aim To compare the observation results of atomic force microscopy(AFM)and scanning electron microscopy(SEM),and to summarize the main problems and solutions of AFM in observing biological macromolecules,using the observa-tion subjects of protein samples purified by our research group.Methods The protein samples were diluted to 15 nmol·L-1 with PBS,fixed on glass slides,silicon wafers,and mica sheets,dried,and made into solid-phase observation samples.SEM sam-ples were plated with platinum before observation.The surface structures of proteins were observed using AFM and SEM,sample heights were calculated,and differences in results were com-pared.Results Protein samples with positive charges tended to shift to the right during observation due to the repulsion of the AFM probe;mica sheets could effectively eliminate the positive charge of proteins to avoid sample movement;PBS provided a stable environment for protein samples,but the crystallization of PBS salts interfered with probe operation and imaging clarity;SEM samples needed to be plated with platinum before observa-tion and could not achieve the precision of AFM.Conclusions Both AFM and SEM can directly observe protein structures in vitro,with AFM providing higher precision results;when protein sample stability permits,ultrapure water is preferred as the sol-vent carrier,and volatile liquids such as ethanol can also serve as solvent carriers.The application of AFM offers a new approach for pharmacological studies on interactions between biological macromolecules.


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