1.Effect of Go-Ichi-Ni-San complex subunit 1 on disease progression and chemotherapy resistance in hepatocellular carcinoma
Yishan HUO ; Dawei LI ; Xiangbing DUAN ; Yuyu MA ; Guojun ZHANG ; Kainan ZHANG ; Xiumin MA
Journal of Clinical Hepatology 2025;41(3):485-492
ObjectiveTo investigate the role and mechanism of Go-Ichi-Ni-San complex subunit 1 (GINS1) in the progression of hepatocellular carcinoma (HCC) and the development of chemotherapy resistance. MethodsThe tumor database GEPIA2 was used to analyze the differential expression of GINS1 between HCC patients and healthy individuals, and pathological tissue samples were collected from 40 HCC patients who were admitted to The Affiliated Tumor Hospital of Xinjiang Medical University and the First Affiliated Hospital of Xinjiang Medical University from May 2017 to January 2021. Immunohistochemical staining was used to measure the difference in the expression of GINS1 between HCC tissue and corresponding adjacent tissue, and the correlation between the expression level of GINS1 and the clinical TNM stage of HCC was analyzed. Western blot was also used to measure the difference in the expression of GINS1 between HCC Huh7/Hep3B/Li-7/MHCC97H cell lines and normal human QSG7701 hepatocytes. The method of lentivirus transfection was used to establish the MHCC97H cell line with stable GINS1 knockdown and its negative control cell line. CCK-8 assay and colony formation assay were used to measure cell proliferative capacity; scratch assay was used to measure cell migration ability; Transwell assay was used to measure cell invasion ability; cells were treated with oxaliplatin to measure their sensitivity to chemotherapy drugs. Nude mice were used to establish a tumor-bearing model and observe the effect of GINS1 knockdown on the growth of HCC in vivo. Western Blot was used to measure the expression levels of the proteins associated with the Notch pathway and the JAK/STAT pathway. The cells were treated with the Notch receptor agonist Jagged-1 to analyze the association between GINS1 and the Notch/JAK/STAT pathway. The independent-samples t test was used for comparison of continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsThe expression of GINS1 was upregulated in HCC patients, HCC tissue, and HCC cell lines (all P<0.05), and the expression level of GINS1 was positively correlated with the clinical TNM stage of HCC (r=0.822, P=0.011). Compared with the negative control cells, the GINS1-knockdown MHCC97H cells showed significant reductions in proliferation, migration, and invasion activities (all P<0.01) and a significantly enhanced sensitivity to oxaliplatin (P<0.01). Compared with the nude mice in the control group, GINS1 knockdown caused significant inhibition of tumor weight and volume in vivo in nude mice (all P<0.001). Compared with the negative control cells, the GINS1-knockdown MHCC97H cells showed significant reductions in the expression levels of Notch1, Notch3, p-JAK2, and p-STAT3 (all P<0.05), while there were no significant differences in the overall expression levels of JAK2 and STAT3 (P>0.05). After Jagged-1 treatment, the GINS1-knockdown MHCC97H cells showed significant increases in proliferation, migration, and invasion activities and a significant reduction in sensitivity to oxaliplatin, as well as significant increases in the levels of p-JAK2 and p-STAT3 (all P<0.05). ConclusionGINS1 is upregulated in HCC and can promote HCC progression and chemotherapy resistance through the Notch/JAK2/STAT3 pathway.
2.A new method for flow cytometry-based detection of ABO antigen expression levels
Yuyu ZHANG ; Xi LIU ; Junhua XIE ; Bin CAO ; Jiewei ZHENG ; Xinyi ZHU ; Zhongying WANG ; Dong XIANG
Chinese Journal of Blood Transfusion 2025;38(5):665-672
Objective: To design and establish a new method for flow cytometry-based detection of commonly observed highly expressed antigens on red blood cells, and to further evaluate the differences and distribution characteristics of antigen expression levels between ABO blood type homozygotes and heterozygotes in healthy individuals. Methods: Residual blood samples after donor blood type identification by Shanghai Blood Center in April 2024 were collected. Among them, samples of 19 homozygous and 19 heterozygous individuals of type A and type B were selected. Then the expression level of ABO antigen on red blood cells were detected using the new method established in this study and the traditional aldehyde fixed red blood cell method. Both methods were tested independently three times and the results were compared. Results: The mean values of the three detection results of the new method was (×10
/RBC): AA homozygous 3.3±0.5, AO heterozygous 2.8±0.3, BB homozygous 3.6±0.3, BO heterozygous 3.1±2.8. The mean values of the three detection results of the aldehyde fixation method were AA homozygous 5.9±0.9, AO heterozygous 5.0±1.4, BB homozygous 3.8±0.6, and BO heterozygous 3.3±0.4. The average antigen distribution of each genotype followed a normal distribution. Comparing the average antigen expression levels of homozygotes and heterozygotes, both methods showed that A/B homozygotes had higher antigen levels than heterozygotes, with AA being 1.17 to 1.18 times that of AO and BB being 1.15 to 1.16 times that of BO. Comparing the inter batch differences in the three test results of two methods, the new method showed no significant difference in the three test results for four genotypes (P>0.05). The aldehyde fixation method showed significant differences in the test results for all three genotypes (P<0.01) except for BB homozygotes (P>0.05). The reliability and reproducibility of the new method were better than those of the traditional aldehyde fixation method. Conclusion: The antigen expression level of ABO homozygotes is higher than that of heterozygotes, and the difference in antigen level between type A homozygotes and heterozygotes is slightly higher than that of type B. The new method is superior to traditional aldolization fixation methods.
3.Differential Analysis on Chemical Composition and Pharmacodynamic Effect Between Combined Decoction and Single Decoction of Famous Classical Formula Huaganjian
Yang WANG ; Gaoju ZHANG ; Ling LI ; Liping CHEN ; Li ZHANG ; Xiao LIU ; Yuyu ZHANG ; Yuan CUI ; Minglong LI ; Chaomei FU ; Xin YAN ; Yuxin HE ; Qin DONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):199-207
ObjectiveThrough qualitatively and quantitatively analysis of the differences in chemical composition between the combined decoction and single decoction of Huaganjian and comparison of their core efficacy, to explore the rationality of the flexible clinical application of Huaganjian compound preparations and single-flavored dispensing granules. MethodsUltra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used to qualitatively analyze the combined decoction and single decoction samples of Huaganjian, and meanwhile, the contents of four index components(geniposide, paeoniflorin, hesperidin and paeonol) were quantitatively analyzed by high performance liquid chromatography(HPLC). Nonalcoholic fatty liver disease(NAFLD) rat model induced by high-fat diet was applied to compare the efficacy of combined decoction and single decoction of Huaganjian. A total of 30 male SD rats were randomly divided into the control group, model group, lovastatin group(1.8 mg·kg-1), combined decoction group(1.26 g·kg-1) and single decoction group(1.18 g·kg-1). After successful modeling, lovastatin group, combined decoction group and single decoction group were given corresponding doses of drugs by intragastric administration every day, and the control group and model group were given equal amounts of normal saline by intragastric administration, after 4 weeks of administration, the serum and liver tissues were collected, and the contents of alanine aminotransferase(ALT), aspartate aminotransferase(AST), total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) in serum of rats were detected, and the liver pathological examination was carried out by hematoxylin-eosin(HE) staining and oil red O staining, so as to compare differences of their efficacy. ResultsSeventy chemical components were initially identified and attributed from the lyophilized powder of the combined decoction and single decoction samples of Huaganjian, and there was no obvious difference in composition between the two. Further quantitative analysis showed that the contents of geniposide, paeoniflorin, hesperidin and paeonol in the combined decoction samples were significantly increased when compared with those of the single decoction samples(P<0.01). The pharmacodynamic results showed that compared with the model group, both the combined and single decoction groups of Huaganjian could improve the liver index of NAFLD rats, reduce the serum levels of AST, ALT, TC, TG and LDL-C, increase the serum level of HDL-C, and ameliorate the pathological changes of liver cell steatosis and fat accumulation. However, there was no significant difference in pharmacodynamic effects between the combined decoction group and the single decoction group. ConclusionThere is no significant difference between the combined decoction and single decoction of Huaganjian in terms of chemical composition, but the contents of the four index components show significantly difference. Both of them can significantly improve the fat accumulation and liver function in NAFLD rats. This study provides a reference basis for the rational clinical application and evaluation of famous classical formula compound preparations and single-flavored dispensing granules.
4.Analysis of Animal Models of Autoimmune Thyroiditis Based on Clinical Characteristics of Traditional Chinese and Western Medicine
Sifeng JIA ; Zhuo ZHANG ; Yuyu DUAN ; Keqiu YAN ; Xinhe ZUO ; Yang LI ; Yong ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(18):235-243
ObjectiveAutoimmune thyroiditis (AIT) is a complex and immune-mediated disorder, with no established treatment protocol. Both Western and traditional Chinese medicine (TCM) focus on the pathogenesis and treatment of AIT. This study evaluated the clinical consistency of existing AIT animal models based on the diagnostic criteria of both Western and TCM, using a novel evaluation method. Additionally, it proposed recommendations and future prospects for improving these models. MethodsA comprehensive literature review was conducted on existing AIT animal models, using databases and the diagnostic criteria of both Western and TCM. Core and accompanying symptoms of these models were scored based on the diagnostic criteria of both Western and TCM, and clinical consistency was assessed. ResultsMice are the primary experimental animals used in AIT modeling. Modeling methods include vaccine immunization, iodine induction, heterologous thyroid antigen immunization, and a combination of high iodine water and antigen immunization. The average consistency of clinical syndromes based on TCM and Western medicine is 40%, 60%, 54%, and 63%, with the highest consistency observed in the combined high iodine water and antigen immunization model. Pathological models based on TCM are less common, with the liver-stagnation-spleen-deficiency rat model showing high clinical consistency. While most models are designed according to Western medical theory, meeting the surface and structural effectiveness criteria of Western medicine. However, there is a lack of fine-tuning and clear differentiation of TCM syndromes. ConclusionCurrent AIT syndrome-disease combination animal models primarily reflect the pathological features of Western medicine, with limited integration of TCM syndromes. Future research should aim to combine the syndrome characteristics of TCM with the pathological features of Western medicine, creating multi-factor and dynamic syndrome-disease models. Such models would better facilitate an experimental platform that conforms to the theories of TCM, providing more comprehensive support and guidance for the pathogenesis and treatment strategies of AIT.
5.Mechanism of Cuscutae Semen-Salviae Miltiorrhizae Radix et Rhizoma in Regulating IL-6/STAT3 Signaling Pathway to Treat Recurrent Spontaneous Abortion Based on Network Pharmacology and Cell Experiment Validation
Haoxian WANG ; Jiuxian LI ; Yan LIU ; Shuo SUN ; Xiangyu LI ; Ke ZHANG ; Yuyu WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):22-30
ObjectiveTo explore the mechanism of action of Cuscutae Semen-Salviae Miltiorrhizae Radix et Rhizoma in the treatment of recurrent spontaneous abortion (RSA) through network pharmacology, molecular docking, and cell experiment verification. MethodsThe Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and UniPort databases were used to screen and organize the active ingredients and corresponding targets of Cuscutae Semen-Salviae Miltiorrhizae Radix et Rhizoma. The potential therapeutic targets of RSA were screened in Online Mendelian Inheritance in Man (OMIM), GeneCards database, DrugBank database, DisGeNET database, and Therapeutic Target Database (TTD). The potential core targets of Cuscutae Semen-Salviae miltiorrhizae Radix et Rhizoma for treating RSA were further screened by constructing a protein-protein interaction (PPI) network and topological analysis. Meanwhile, the Database for Annotation, Visualization and Integrated Discovery (DAVID) was chosen to perform enrichment analysis on intersection targets. On this basis, AutoDock software was used for molecular docking, and the data were imported into PyMOL software for visualization and composition. Finally, the cell counting kit-8 (CCK-8) experiment, Transwell cell invasion assay, and Western blot were used to detect the effects of serum containing Cuscutae Semen-Salviae miltiorrhizae Radix et Rhizoma on HTR-8/SVneo cells and observe the effects on the interleukin (IL)-6/signal transducer and activator of transcription 3(STAT3) signaling pathway and related proteins. ResultsThrough network pharmacology analysis, a total of 69 active ingredients, 73 potential therapeutic targets, and 17 core targets, including IL-6, IL-10, and STAT3, were collected. The 73 common targets were enriched in 614 Gene Ontology (GO) entries and 57 Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways. The molecular docking results indicated that IL-6 and STAT3 had good binding ability with the main active ingredients, including matrine, cryptotanshinone, and tanshinone ⅡA. The cell experiment results showed that, compared with those of the control group, after 24 hours of treatment with the drug-containing serum, the survival and invasion rates of HTR-8/SVneo cells were significantly increased (P<0.05), and the expression of IL-6/STAT3 signaling pathway and related proteins IL-10 and c-Myc was significantly elevated (P<0.05). Moreover, the trend of action in the drug-containing serum group was consistent with that of pathway agonists. ConclusionCuscutae Semen-Salviae miltiorrhizae Radix et Rhizoma may enhance the survival rate and invasive activity of HTR-8/SVneo cells to further prevent and treat RSA by activating the IL-6/STAT3 signaling pathway and upregulating the expression of downstream factors IL-10 and c-Myc in the pathway.
6.Clinical Application and Pharmacological Mechanism of Sishenwan in Treatment of Ulcerative Colitis: A Review
Keqiu YAN ; Xiaoyu ZHANG ; Sifeng JIA ; Yuyu DUAN ; Zixing QIAN ; Yifan CAI ; Junyi SHEN ; Wenjie XIAO ; Xinkun BAO ; Guangjun SUN ; Aizhen LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):261-270
Ulcerative colitis (UC), a chronic, non-specific inflammatory bowel disease with typical symptoms such as abdominal pain, diarrhea, and bloody stools, demonstrates a high relapse rate and difficulty in curing. Sishenwan, first recorded in Internal Medicine Abstract (Nei Ke Zhai Yao), are a classic prescription for treating diarrhea caused by deficiency of the spleen and kidney Yang. The core therapeutic principle of Sishenwan is warming and tonifying the spleen and kidney, and astringing the intestine and stopping diarrhea. In recent years, Sishenwan have demonstrated distinct advantages in the clinical treatment of UC. The pathogenesis of UC involves multiple factors, including immune dysregulation and gut microbiota imbalance. Although Western medicine is effective in the short term, its side effects, high relapse rate, and resistance associated with long-term use pose substantial challenges. Sishenwan have shown excellent clinical outcomes in the treatment of UC due to deficiency of the spleen and kidney Yang. Modern clinical studies indicate that Sishenwan, used alone or in combination with Western medicine or other Chinese medicine compound prescriptions, significantly improve the clinical efficacy in treating UC due to deficiency of the spleen and kidney Yang. Sishenwan effectively alleviate core symptoms such as mucus, pus, and blood in stools, and persistent abdominal pain, reduce Mayo scores and the relapse rate, and improve patients' quality of life. Research on the material basis reveals that Sishenwan contain multiple active ingredients such as psoralen, isopsoralen, and evodiamine. Mechanism studies indicate that Sishenwan inhibit the inflammatory cascade reactions by regulating the signal network through multiple targets. Sishenwan regulate cellular immunity and restore intestinal immune homeostasis. At the microecological level, Sishenwan promote the intestinal barrier repair through the "microbiota-metabolism-immunity" axis. The current research still needs to be deepened in aspects such as the mining of specific biomarkers for syndromes and the exploration of the collaborative mechanism of traditional Chinese and Western medicine. In the future, a full-chain system covering syndrome differentiation, targeting, and monitoring needs to be constructed for promoting the paradigm transformation of Sishenwan into precision drugs. This review systematically explains the treatment mechanism of Sishenwan regarding the combination of disease and syndrome and its multi-target regulatory characteristics, providing a theoretical basis and transformation direction for the treatment of UC with integrated traditional Chinese and Western medicine.
7.Clinical Application and Pharmacological Mechanism of Sishenwan in Treatment of Ulcerative Colitis: A Review
Keqiu YAN ; Xiaoyu ZHANG ; Sifeng JIA ; Yuyu DUAN ; Zixing QIAN ; Yifan CAI ; Junyi SHEN ; Wenjie XIAO ; Xinkun BAO ; Guangjun SUN ; Aizhen LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):261-270
Ulcerative colitis (UC), a chronic, non-specific inflammatory bowel disease with typical symptoms such as abdominal pain, diarrhea, and bloody stools, demonstrates a high relapse rate and difficulty in curing. Sishenwan, first recorded in Internal Medicine Abstract (Nei Ke Zhai Yao), are a classic prescription for treating diarrhea caused by deficiency of the spleen and kidney Yang. The core therapeutic principle of Sishenwan is warming and tonifying the spleen and kidney, and astringing the intestine and stopping diarrhea. In recent years, Sishenwan have demonstrated distinct advantages in the clinical treatment of UC. The pathogenesis of UC involves multiple factors, including immune dysregulation and gut microbiota imbalance. Although Western medicine is effective in the short term, its side effects, high relapse rate, and resistance associated with long-term use pose substantial challenges. Sishenwan have shown excellent clinical outcomes in the treatment of UC due to deficiency of the spleen and kidney Yang. Modern clinical studies indicate that Sishenwan, used alone or in combination with Western medicine or other Chinese medicine compound prescriptions, significantly improve the clinical efficacy in treating UC due to deficiency of the spleen and kidney Yang. Sishenwan effectively alleviate core symptoms such as mucus, pus, and blood in stools, and persistent abdominal pain, reduce Mayo scores and the relapse rate, and improve patients' quality of life. Research on the material basis reveals that Sishenwan contain multiple active ingredients such as psoralen, isopsoralen, and evodiamine. Mechanism studies indicate that Sishenwan inhibit the inflammatory cascade reactions by regulating the signal network through multiple targets. Sishenwan regulate cellular immunity and restore intestinal immune homeostasis. At the microecological level, Sishenwan promote the intestinal barrier repair through the "microbiota-metabolism-immunity" axis. The current research still needs to be deepened in aspects such as the mining of specific biomarkers for syndromes and the exploration of the collaborative mechanism of traditional Chinese and Western medicine. In the future, a full-chain system covering syndrome differentiation, targeting, and monitoring needs to be constructed for promoting the paradigm transformation of Sishenwan into precision drugs. This review systematically explains the treatment mechanism of Sishenwan regarding the combination of disease and syndrome and its multi-target regulatory characteristics, providing a theoretical basis and transformation direction for the treatment of UC with integrated traditional Chinese and Western medicine.
8.Research on the application rules of aromatic Chinese herbs in the prevention and treatment of warm diseases
Chun WANG ; Linyuan WANG ; Jianjun ZHANG ; Linlin XIU ; Yuyu HE ; Yuxin JIA ; Weican LIANG ; Yi LI ; Yinming ZHAO
Journal of Beijing University of Traditional Chinese Medicine 2025;48(4):451-458
Traditional Chinese medicine (TCM) has historically played a pivotal role in the prevention and treatment of warm diseases, establishing a comprehensive theoretical framework that underpins its practices. The distinctive and indispensable contributions of aromatic Chinese herbs in dispelling harmful influences and mitigating the spread of these diseases are well recognized; however, further investigation is warranted to elucidate their systematic properties and regularities, and the theory of aromatic Chinese herbs in preventing and treating warm diseases still needs to be comprehensively summarized. This study employs the principles rooted in TCM, with particular emphasis on the framework for warm diseases. An analysis of the disease mechanisms, transmission dynamics, and preventive strategies is conducted during the early stage of infection, throughout the course of the disease, and in the post-illness phase. Furthermore, the characteristics and applications of aromatic Chinese herbs are integrated with insights drawn from modern pharmacological research to explore their specific roles in the prevention and management of warm diseases. The utilization of aromatic Chinese herbs manifests in a variety of therapeutic effects: aromatic medicinals purging filth and dispelling pathogens for preventing epidemic disease, aromatic medicinals regulation for relieving superficies syndrome and dispersing evils, aromatic medicinals ventilation the lung to relieve cough and asthma, aromatic medicinals resolving the dampness to awaken the spleen and stomach, aromatic medicinals opening the orifices to restore consciousness, aromatic and pungent medicinals to regulate qi, aromatic medicinals dredging the vessels to activate blood circulation and dissipate blood stasis, and aromatic medicinals clearing latent heat from the yin level. These properties facilitate tailored approaches to address the diverse manifestations of warm diseases and their associated symptoms, providing clear guidance for clinical application to achieve pre-disease prevention, active disease treatment, complication prevention, and post-recovery relapse avoidance. The use of aromatic Chinese herbs in preventing and treating warm diseases demonstrates theoretical, practical, systematic, and regular characteristics. The theory of the properties of aromatic Chinese herbs has been expanded and sublimated in clinical practice, and its scientific connotation has been expounded in modern research. Under the guidance of the theory of treatment based on syndrome differentiation, and by taking into account the distinct stages and pathologies of warm diseases, the rational selection of aromatic Chinese herbs can improve the clinical efficacy.
9.Determination of Traditional Chinese Medicine Syndrome of Rat Model of Follicular Dysplasia Induced by Tripterygium Glycosides Based on Prescriptions via AMPK/mTOR/HIF-1/VEGF Pathway
Yingying HU ; Xuejuan LIN ; Zushun ZHUO ; Shanshan DING ; Mengting ZHANG ; Guorong LIN ; Long ZHU ; Yuyu XIE ; Yan XIONG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):46-54
ObjectiveTo determine the syndrome of a rat model of follicular dysplasia induced by Tripterygium glycosides based on prescriptions and investigate the mechanism of traditional Chinese medicine intervention via the adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)/hypoxia-inducible factor-1 (HIF-1)/vascular endothelial growth factor (VEGF) pathway. MethodForty-eight rats with regular estrous cycles were randomly assigned into a normal group (n=8) and a modeling group (n=40). The rats in the modeling group were administrated with Tripterygium glycoside suspension (75 mL·kg-1) by gavage for 30 days. The modeled rats were assigned into model, Siwutang (3.69 g·kg-1), Youguiyin (3.11 g·kg-1), Zuoguiyin (7.29 g·kg-1), and Guishenwan (10.35 g·kg-1) groups, with 8 rats in each group. The drug intervention lasted for 14 days. The changes of estrous cycle were detected by Pap staining, and a stereoscope was used to observe the morphology of the ovarian tissue. Hematoxylin-eosin staining was employed to observe the pathological changes and follicle count in the ovarian tissue. Enzyme-related immunosorbent assay (ELISA) was used to measure the levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) in the serum. Real-time fluorescence quantitative polymerase chain reaction and Western blot were employed to determine the mRNA and protein levels, respectively, of AMPK, mTOR, HIF-1, and VEGF in the ovarian tissue. ResultCompared with the normal group, the model group had a disordered estrous cycle, reduced secondary and mature follicles, increased atretic follicles, elevated FSH and LH levels, lowered E2 level, up-regulated mRNA and protein levels of AMPK, and down-regulated mRNA and protein levels of mTOR, HIF-1, and VEGF (P<0.01). Compared with the model group, Guishenwan increased secondary and mature follicles, decreased atretic follicles, lowered the FSH and LH levels, elevated the E2 level, down-regulated the mRNA and protein levels of AMPK, and up-regulated the mRNA and protein levels of mTOR, HIF-1, and VEGF (P<0.01). Compared with Guishenwan group, Siwutang, Youguiyin, and Zuoguiyin decreased mature follicles, increased atretic follicles (P<0.01), elevated the LH (P<0.01) and FSH (P<0.05) levels, and lowered the E2 level (P<0.05). In addition, Youguiyin up-regulated the protein level of AMPK (P<0.05) and down-regulated the mRNA levels of mTOR and HIF-1 (P<0.01) as well as the mRNA and protein levels of VEGF (P<0.01). Siwutang down-regulated the mRNA levels of mTOR and HIF-1 as well as the mRNA and protein levels of VEGF (P<0.05). Zuoguiyin down-regulated the mRNA level of mTOR and the protein and mRNA levels of VEGF (P<0.05). ConclusionGuishenwan may improve the ovarian function and promote follicle maturation in a rat model of follicular dysplasia by inhibiting the AMPK/mTOR/HIF-1/VEGF pathway, with the therapeutic effect superior to Zuoguiyin, Youguiyin, and Siwutang. It was hypothesized that this model presented the syndrome of kidney-essence deficiency.
10.Effects of targeting modification on intracellular transportation of PEG-PCL micelles in human cervical cancer cells
Jinjin YANG ; Qinghua YU ; Lingbo YU ; Yadong ZHANG ; Dongqin LIANG ; Yuyu SUN ; Huiyun WANG ; Yanan CUI
China Pharmacy 2024;35(12):1431-1436
OBJECTIVE To study the effects of transferrin-targeting peptide T7 (7pep) on intracellular transportation of polyethylene glycol-polycaprolactone (PEG-PCL) micelles in human cervical cancer HeLa cells. METHODS Using coumarin-6 (C6) as fluorescent indicator probe, both coumarin-6 (C6)-loaded PEG-PCL (PEG-PCL-C6) micelles and 7pep-modified PEG- PCL (7pep-PEG-PCL-C6) micelles were prepared by film-dispersion method. The particle size, polydispersity index and appearance morphology were compared between two types of micelles; the real-time uptake of two types of micelles by HeLa cells was compared, and the colocalization of two types of micelles with early endosomes (EE), endocytic recycling compartments (ERC) and late endosomes (LE) after entry into the cells was observed. RESULTS The particle sizes of PEG-PCL-C6 and 7pep-PEG-PCL- C6 micelles were(75.0±2.3)and(82.0±1.5)nm; the polymer dispersity indexes were 0.17±0.20 and 0.17±0.32, respectively, with a regular spherical appearance. The colocalization results showed that entry speed and amount of 7pep-PEG-PCL-C6 micelles were significantly faster/more than those of PEG-PCL-C6 micelles. 7pep-PEG-PCL-C6 micelles entered EE faster than PEG-PCL-C6 micelles, while PEG-PCL-C6 micelles entered ERC at a faster rate than 7pep-PEG-PCL-C6 micelles, and both PEG-PCL-C6 micelles and 7pep-PEG-PCL-C6 micelles tended to accumulate gradually in LE; Pearson coefficient, signal overlap ratio, and colocalization ratio of 7pep-PEG-PCL-C6 micelles with LE were significantly lower 60 minutes after entering the cell than those 30 minutes after entering the cell (P<0.05 or P<0.01). CONCLUSIONS Targeting 7pep modification can increase the entry speed and amount of PEG-PCL-C6 micelles, and also alter their intracellular transportation behavior.


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