1.Quality Evaluation of Naomaili Granules Based on Multi-component Content Determination and Fingerprint and Screening of Its Anti-neuroinflammatory Substance Basis
Ya WANG ; Yanan KANG ; Bo LIU ; Zimo WANG ; Xuan ZHANG ; Wei LAN ; Wen ZHANG ; Lu YANG ; Yi SUN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):170-178
ObjectiveTo establish an ultra-performance liquid fingerprint and multi-components determination method for Naomaili granules. To evaluate the quality of different batches by chemometrics, and the anti-neuroinflammatory effects of water extract and main components of Naomaili granules were tested in vitro. MethodsThe similarity and common peaks of 27 batches of Naomaili granules were evaluated by using Ultra performance liquid chromatography (UPLC) fingerprint detection. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technology was used to determine the content of the index components in Naomaili granules and to evaluate the quality of different batches of Naomaili granules by chemometrics. LPS-induced BV-2 cell inflammation model was used to investigate the anti-neuroinflammatory effects of the water extract and main components of Naomaili granules. ResultsThe similarity of fingerprints of 27 batches of samples was > 0.90. A total of 32 common peaks were calibrated, and 23 of them were identified and assigned. In 27 batches of Naomaili granules, the mass fractions of 14 components that were stachydrine hydrochloride, leonurine hydrochloride, calycosin-7-O-glucoside, calycosin,tanshinoneⅠ, cryptotanshinone, tanshinoneⅡA, ginsenoside Rb1, notoginsenoside R1, ginsenoside Rg1, paeoniflorin, albiflorin, lactiflorin, and salvianolic acid B were found to be 2.902-3.498, 0.233-0.343, 0.111-0.301, 0.07-0.152, 0.136-0.228, 0.195-0.390, 0.324-0.482, 1.056-1.435, 0.271-0.397, 1.318-1.649, 3.038-4.059, 2.263-3.455, 0.152-0.232, 2.931-3.991 mg∙g-1, respectively. Multivariate statistical analysis showed that paeoniflorin, ginsenoside Rg1, ginsenoside Rb1 and staphylline hydrochloride were quality difference markers to control the stability of the preparation. The results of bioactive experiment showed that the water extract of Naomaili granules and the eight main components with high content in the prescription had a dose-dependent inhibitory effect on the release of NO in the cell supernatant. Among them, salvianolic acid B and ginsenoside Rb1 had strong anti-inflammatory activity, with IC50 values of (36.11±0.15) mg∙L-1 and (27.24±0.54) mg∙L-1, respectively. ConclusionThe quality evaluation method of Naomaili granules established in this study was accurate and reproducible. Four quality difference markers were screened out, and eight key pharmacodynamic substances of Naomaili granules against neuroinflammation were screened out by in vitro cell experiments.
2.Molecular Mechanism of Astragali Radix and Hedyotis diffusa in Regulating LINC01134-CTCF-p21 Axis to Inhibit Lung Adenocarcinoma Proliferation
Haipeng SUN ; He ZHUANG ; Xue LIU ; Siyuan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):131-138
ObjectiveTo explore the interaction and competitive binding of Homo sapiens long intergenic non-protein-coding RNA 1134 (LINC01134) to CCCTC-binding factor CTCF, affecting the transcription of cyclin-dependent kinase inhibitor (p21) and influencing the proliferation of A549 cells, in order to investigate the possible mechanism of Astragali Radix and Hedyotis diffusa (A-H) in inhibiting A549 proliferation by regulating this axis. MethodsRNA-binding protein immunoprecipitation (RIP) assays were conducted to examine the interaction between LINC01134 and CTCF, and chromatin immunoprecipitation (ChIP) assays were used to study the effect of LINC01134 overexpression on the interaction between CTCF and p21. Stable A549 cell lines (oe-NC and oe-LINC01134) were established using lentiviral transfection, and each group was treated with 10% A-H drug-containing serum. Real-time PCR and Western blot analyses were performed to detect the effects of A-H on the expression of LINC01134, CTCF, and p21 in A549 cells. Cell counting kit-8 (CCK-8) and colony formation assays were used to assess the effects of A-H on A549 cell proliferation via LINC01134. Flow cytometry was employed to evaluate the effects of A-H on the A549 cell cycle through LINC01134, and Western blot was used to detect changes in cell cycle proteins. ResultsCompared with the IgG group, the oe-CTCF group showed a significantly increased abundance of LINC01134 aggregates (P0.01). Compared with the oe-Vector group, p21 abundance in CTCF complexes was significantly reduced in the oe-LINC01134 group (P0.01). Compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed the expression of LINC01134 and p21 (P0.05), but had no significant regulatory effect on CTCF. Compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed cell viability at 72 h (P0.05), inhibited malignant proliferation (P0.05), and reversed the proportions of cells in the G0/G1 and S phases (P0.01). Furthermore, compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed the expression of Cyclin D1, CDK4, Cyclin E, CDK2, phosphorylated retinoblastoma protein (p-Rb), and E2F transcription factor 3 (E2F3) (P0.01). ConclusionA-H regulates the LINC01134-CTCF-p21 axis to block the G1/S phase transition of A549 cell cycle, accelerate cellular senescence, and inhibit malignant proliferation.
3.Molecular Mechanism of Astragali Radix and Hedyotis diffusa in Regulating LINC01134-CTCF-p21 Axis to Inhibit Lung Adenocarcinoma Proliferation
Haipeng SUN ; He ZHUANG ; Xue LIU ; Siyuan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):131-138
ObjectiveTo explore the interaction and competitive binding of Homo sapiens long intergenic non-protein-coding RNA 1134 (LINC01134) to CCCTC-binding factor CTCF, affecting the transcription of cyclin-dependent kinase inhibitor (p21) and influencing the proliferation of A549 cells, in order to investigate the possible mechanism of Astragali Radix and Hedyotis diffusa (A-H) in inhibiting A549 proliferation by regulating this axis. MethodsRNA-binding protein immunoprecipitation (RIP) assays were conducted to examine the interaction between LINC01134 and CTCF, and chromatin immunoprecipitation (ChIP) assays were used to study the effect of LINC01134 overexpression on the interaction between CTCF and p21. Stable A549 cell lines (oe-NC and oe-LINC01134) were established using lentiviral transfection, and each group was treated with 10% A-H drug-containing serum. Real-time PCR and Western blot analyses were performed to detect the effects of A-H on the expression of LINC01134, CTCF, and p21 in A549 cells. Cell counting kit-8 (CCK-8) and colony formation assays were used to assess the effects of A-H on A549 cell proliferation via LINC01134. Flow cytometry was employed to evaluate the effects of A-H on the A549 cell cycle through LINC01134, and Western blot was used to detect changes in cell cycle proteins. ResultsCompared with the IgG group, the oe-CTCF group showed a significantly increased abundance of LINC01134 aggregates (P0.01). Compared with the oe-Vector group, p21 abundance in CTCF complexes was significantly reduced in the oe-LINC01134 group (P0.01). Compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed the expression of LINC01134 and p21 (P0.05), but had no significant regulatory effect on CTCF. Compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed cell viability at 72 h (P0.05), inhibited malignant proliferation (P0.05), and reversed the proportions of cells in the G0/G1 and S phases (P0.01). Furthermore, compared with the 10% blank + oe-LINC01134 group, the 10% A-H + oe-LINC01134 group reversed the expression of Cyclin D1, CDK4, Cyclin E, CDK2, phosphorylated retinoblastoma protein (p-Rb), and E2F transcription factor 3 (E2F3) (P0.01). ConclusionA-H regulates the LINC01134-CTCF-p21 axis to block the G1/S phase transition of A549 cell cycle, accelerate cellular senescence, and inhibit malignant proliferation.
4.Pathogenesis and Prevention Strategies of Hypercoagulable State in Malignant Tumors Based on the Theory of "Sweet-Flavored Medicinals Retaining and Restoring Body Fluid"
Yong WANG ; Zixuan CHENG ; Weiyang KONG ; Yuwei SUN ; Yunxuan SHI ; Ruyu QIN ; Zhaidong LIU
Journal of Traditional Chinese Medicine 2026;67(1):26-30
Based on the theory of "sweet-flavored medicinals retaining and restoring body fluid", this paper proposed that the core pathogenesis of hypercoagulable state in malignant tumors is qi deficiency and fluid consumption, blood stasis and vessels stagnation, which evolves dynamically according to the pattern "qi deficiency → fluid consumption → blood stasis". Accordingly, a staged treatment system is established with the general principle of "fortifying the middle jiao, restoring fluid and activating blood circulation". In the initial stage, invigorating the spleen and boosting qi to generate body fluid, targeting the onset of middle jiao deficiency and body fluid consumption; in the middle stage, nourishing yin and unblocking collaterals to facilitate body fluid circulation, addressing the disorder of body fluid transportation and collateral injury caused by internal dryness; in the late stage, consolidating yin and resolving blood stasis to retain body fluid, resolving yin impairment, fluid exhaustion, and binding of stasis and toxin. By regulating body fluid metabolism to improve the hypercoagulable state, this system is intended to provide insights for the prevention and treatment of hypercoagulable state in malignant tumors with traditional Chinese medicine.
5.Effect of Qingfei Shenshi Decoction (清肺渗湿汤) Combined with Western Medicine on Clinical Effectiveness and Immune Function for Patients with Bronchial Asthma of Heat Wheezing Syndrome
Ying SUN ; Haibo HU ; Na LIU ; Fengchan WANG ; Jinbao ZONG ; Ping HAN ; Peng LI ; Guojing ZHAO ; Haoran WANG ; Xuechao LU
Journal of Traditional Chinese Medicine 2026;67(1):38-44
ObjectiveTo observe the clinical effectiveness and safety of Qingfei Shenshi Decoction (清肺渗湿汤) combined with western medicine for patients with bronchial asthma of heat wheezing syndrome, and to explore its potential mechanism of action. MethodsEighty-six participants with bronchial asthma of heat wheezing syndrome were randomly divided into treatment group and control group, each group with 43 participants. The control group received conventional western medicine, and the treatment group was additionally administered Qingfei Shenshi Decoction orally on the basis of the control group, 1 dose per day. Both groups were treated for 14 days. The primary outcome measure was clinical effectiveness; secondary outcome measures included traditional Chinese medicine (TCM) syndrome score, asthma control test (ACT) score, pulmonary function indices such as forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), peak expiratory flow (PEF), serum inflammatory factor levels including interleukin-4 (IL-4), tumour necrosis factor-α (TNF-α), and high-sensitivity C-reactive protein (hs-CRP), and immune function indices including CD3+, CD4+, CD8+, CD4+/CD8+. All outcome measures were evaluated before and after treatment. Vital signs were monitored, and electrocardiography, blood routine, urine routine, liver function, and renal function tests were performed before and after treatment. Adverse events and reactions during the study were recorded. ResultsA total of 80 patients completed the trial with 40 in each group. The total clinical effective rate of the treatment group was 97.5% (39/40), which was significantly higher than that of the control group (85.0%, 34/40, P<0.05). After treatment, both groups showed decreased TCM syndrome scores, IL-4, TNF-α, hs-CRP, and CD8+ levels, as well as increased ACT scores, CD3+, CD4+, CD4+/CD8+, FEV1, FVC, and PEF levels (P<0.05 or P<0.01). Moreover, the improvements in these indices were more significant in the treatment group than in the control group (P<0.05 or P<0.01). No significant abnormalities in safety indicators were observed in either group, and no adverse events or reactions occurred. ConclusionQingfei Shenshi Decoction combined with conventional western medicine for patients with bronchial asthma of heat wheezing syndrome can effectively improve the clinical symptoms, pulmonary function, and clinical effectiveness, with good safety. Its mechanism may be related to reducing inflammatory factor levels and regulating T lymphocyte subsets to improve immune function.
6.Research progress on antibody-drug conjugates in the treatment of triple-negative breast cancer
Danna LIU ; Shuangshuang SONG ; Lu CHEN ; Yongqiang SUN ; Bo SUN ; Hanli ZHOU ; Xiaoli ZHAO ; Tiandong KONG
China Pharmacy 2026;37(1):124-129
Antibody-drug conjugates (ADCs) are a novel class of anti-tumor agents composed of a targeted monoclonal antibody, a cytotoxic drug, and a linker connecting the two. They combine the high specificity of antibodies with the potent cytotoxicity of chemotherapeutic agents. Triple-negative breast cancer (TNBC) is characterized by high aggressiveness, elevated risks of recurrence and metastasis, and poor prognosis, largely due to the lack of effective therapeutic targets. This review summarizes the research progress of ADCs in the treatment of TNBC. It has been found that ADCs targeting human epidermal growth factor receptor 2 (such as trastuzumab deruxtecan), trophoblast cell surface antigen 2 (such as sacituzumab govitecan and datopotamab deruxtecan), zinc transporter LIV-1 (such as ladiratuzumab vedotin), HER-3 (such as patritumab deruxtecan), epidermal growth factor receptor (such as AVID100), and glycoprotein non-metastatic melanoma protein B (such as glembatumumab vedotin) have all demonstrated promising therapeutic effects against TNBC. Despite challenges including acquired resistance and treatment-related toxicities, ADCs are undoubtedly reshaping the therapeutic landscape for TNBC and are expected to occupy a more central position in TNBC treatment in the future.
7.Fabrication and evaluation of an inositol hexaphosphate-zinc hydrogel with dual capabilities of self-mineralization and osteoinduction
LIU Mingyi ; MIAO Xiaoyu ; CAI Yunfan ; WANG Yan ; SUN Xiaotang ; KANG Jingrui ; ZHAO Yao ; NIU Lina
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(1):29-40
Objective:
To fabricate a hydrogel loaded with inositol hexaphosphate-zinc and preliminarily evaluate its performance in self-mineralization and osteoinduction, thereby providing a theoretical basis for the development of bone regeneration materials.
Methods:
The hydrogel framework (designated DF0) was formed by copolymerizing methacryloyloxyethyltrimethylammonium chloride and four-armed poly(ethylene glycol) acrylate, followed by sequentially loading inositol hexaphosphate anions via electrostatic interaction and zinc ions via chelation. The hydrogel loaded only with inositol hexaphosphate anions was named DF1, while the co-loaded hydrogel was named DF2. The self-mineralization efficacy of the DF0 , DF1 and DF2 hydrogels was characterized using scanning electron microscopy, transmission electron microscopy (TEM), energy dispersive spectroscopy (EDS), and selected area electron diffraction (SAED). The biocompatibility was assessed via live/dead cell staining and a CCK-8 assay. The osteoinductive capacity of the DF0 , DF1 and DF2 hydrogels on MC3T3-E1 cells was assessed via alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining. In the aforementioned cell experiments, cells cultured in standard medium served as the control group
Results:
The DF0, DF1, and DF2 hydrogels were successfully synthesized. Notably, DF1 and DF2 exhibited distinct self-mineralization within 6 days. Results from TEM, EDS, and SAED confirmed that the mineralization products were amorphous calcium phosphate in group DF1, and amorphous calciumzinc phosphate in group DF2. Biocompatibility tests revealed that none of the hydrogels (DF0, DF1, and DF2) adversely affected cell viability or proliferation. In osteogenic induction experiments, both ALP and ARS staining were intensified in the DF1 and DF2 groups, with the most profound staining observed in the DF2 group.
Conclusion
The developed inositol hexaphosphate-zinc hydrogel (DF2) demonstrates the dual capacity to generate calcium-phosphate compounds through self-mineralization while exhibiting excellent osteoinductive properties. This biocompatible, dual-promoting osteogenic hydrogel presents a novel strategy for bone regeneration.
8.Pharmacokinetic Analysis of Ziyuglycoside Ⅰ in Normal and Acute Kidney Injury Rats
Yunhui ZHANG ; Yanli LIU ; Qiongming XU ; Shuding SUN ; Hongjin ZHU ; Di ZHAO ; Suxiang FENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):203-210
ObjectiveBased on ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap-linear ion-trap mass spectrometry(UPLC-Orbitrap Fusion Lumos Tribrid-MS), the plasma concentration of ziyuglycoside Ⅰ was determined at different time points after oral administration, and its pharmacokinetic characteristics in normal rats and rats with acute kidney injury were compared. MethodsRats were randomly divided into normal group and model group, the model group received intraperitoneal cisplatin(10 mg·kg-1) to establish the acute kidney injury model, the normal group was given the same volume of saline. After successful modeling, rats in the normal and model groups were randomly divided into the normal low, medium and high dose groups(2.5, 5, 7.5 mg·kg-1) and the model low, medium and high dose groups(2.5, 5, 7.5 mg·kg-1), 6 rats in each group, and the plasma was collected at different time points after receiving the corresponding dose of ziyuglycoside Ⅰ. Then, the concentration of ziyuglycoside Ⅰ in rat plasma was determined by UPLC-Orbitrap Fusion Lumos Tribrid-MS, and the drug-time curve was poltted. The pharmacokinetic parameters were calculated by Kinetica 5.1 software, and the differences in pharmacokinetic parameters between different administration groups were compared by independent sample t-test with SPSS 22.0. ResultsThe pharmacokinetic results showed that after receiving the different doses of ziyuglycoside Ⅰ, its concentration increased first and then decreased, and all of them reached the maximum plasma concentration at about 0.5 h. The area under the curve(AUC0-t) and mean retention time(MRT0-t) of normal and model rats increased with the increased dose, and the clearance(CL) decreased with the increasing dose. Compared with the normal group, the AUC0-t was significantly increased(P<0.01), peak concentration(Cmax) and CL decreased in model rats at different doses, indicating that the physiological state of the rats could affect the absorption and elimination of ziyuglycoside Ⅰ in vivo. ConclusionThe pharmacokinetic characteristics of ziyuglycoside Ⅰ are quite different in normal rats and acute kidney injury model rats, which may be due to the change of the body environment in the pathological state, then lead to changes in absorption and metabolic processes.
9.Clinical Diseases Responding Specially to TCM Treatment: Psoriasis
Liu LIU ; Xiaoying SUN ; Mei MO ; Yaqiong ZHOU ; Bin LI ; Xiaoxiao ZHANG ; Xin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):260-268
Psoriasis is a common chronic inflammatory systemic disease in dermatology. Its high prevalence, recurrence rate, and numerous comorbidities impose a significant physical and mental burden on patients. With the continuous advancement of modern medicine, the emergence of biological agents has improved clinical efficacy, making it possible to overcome psoriasis, in addition to classical treatments. However, in clinical practice, adverse reactions, drug resistance, recurrence rates, and immune drift cannot be ignored. Traditional Chinese medicine (TCM) has a history of thousands of years in treating psoriasis, demonstrating good efficacy, high safety, and a low recurrence rate, but a standardized management system is lacking. Therefore, the 25th Clinical Diseases Responding Specially to TCM Treatment Series (Psoriasis) Youth Salon, hosted by the Chinese Association of Chinese Medicine and organized by the Youth Committee of the Chinese Association of Chinese Medicine, invited 29 experts and scholars from TCM, Western medicine, and interdisciplinary fields to actively discuss the "Advantages, Challenges, and Clinical Transformation of TCM and Western Medicine in the Diagnosis and Treatment of Psoriasis". The experts at the meeting concluded that the advantages of TCM in the treatment of psoriasis are as follows. Firstly, in the TCM-led treatment plan, TCM's understanding of psoriasis follows the principle of combining the differentiation of disease and syndrome. This approach distinguishes the basic contradiction from the current main contradiction and enables a clear grasp of the dynamic process of psoriasis development. Based on the system of syndrome differentiation and treatment, TCM intervention is applied to address the current main contradiction, and the optimal TCM treatment plan is formulated by combining internal and external treatments. Adhering to the principle of "what is visible outside must be addressed inside", TCM can prevent and treat psoriasis comorbidities early by differentiating syndrome types. Secondly, in the integrated TCM and Western medicine treatment plan, the combination of both methods not only enhances efficacy but also reduces the adverse reactions of immunosuppressants and biological agents, lowering the recurrence rate. This conference provides a reference for the diagnosis and treatment of psoriasis using TCM and integrated TCM and Western medicine, opening up new ideas for clinical and basic research and guiding future research directions.
10.Exploration on factors influencing HLA-C molecular expression level by flow cytometry
Yunan LI ; Renhui JIANG ; Siqi CAI ; Jie LIU ; Zhihui DENG
Chinese Journal of Blood Transfusion 2025;38(1):79-84
[Objective] To investigate the factors influencing the detection of HLA-C expression by flow cytometry. [Methods] A total of 12 hematopoietic stem cell suspension samples from peripheral hematopoietic stem cell volunteer donors were randomly collected after CD34+ cell counting detection. The influence of detecting different number of nucleated cell (500 000, 50 000 and 5 000), sequential order of red blood cell lysis and antibody incubation, and the HLA-C antibody with varied remaining time from the expiration date on the detection results of HLA-C expression by flow cytometry were investigated, respectively. The significance of differences between different groups was analyzed through Student t test. [Results] There was no significant difference in the proportion of HLA-C positive cells and mean fluorescence intensity (MFI) among the three groups with different nucleated cell numbers detected (500 000, 50 000 and 5 000) (P>0.05). The sequential order of red blood cell lysis and antibody incubation had no influence on the proportion of HLA-C positive cells (P>0.05), but HLA-C MFI value was significantly lower when antibody incubation was performed after red blood cell lysis than that when antibody incubation was performed before red blood cell lysis (P<0.05). The proportion of HLA-C positive cells and MFI value detected by HLA-C antibody remaining 24 months from the expiration date were significantly higher than those detected by HLA-C antibody remaining only 5 months from the expiration date (P<0.05). [Conclusion] The present study has investigated the factors of influencing HLA-C expression level by flow cytometry, the results have important reference and application value for standardizing the experimental operation of HLA-C expression and improving the accuracy and comparability of detection results.


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