1.The Role of Lysosomal Dysfunction in Hepatocellular Carcinoma: From Pathogenesis to Targeted Therapies
Yue-Yan WU ; Xin CHEN ; Ce-Fan ZHOU ; Jing-Feng TANG ; Rui ZHANG
Progress in Biochemistry and Biophysics 2026;53(3):609-622
Hepatocellular carcinoma (HCC) is a lethal cancer with high morbidity rates worldwide. It is a major threat to public health in China, due to the combination of known and new risk factors, such as endemic hepatitis B virus (HBV), dietary aflatoxin exposure, and the occurrence of metabolic dysfunction-associated steatotic liver disease (MASLD). Although many methods for surveillance and multimodal therapies, such as surgery, local ablation, transarterial therapy, and new systemic agents, have been available, the survival rates of HCC remains poor. They have very limited durable responses, long post-treatment recurrence rates, and high resistance to treatment. This reflects an imperfect picture of the biological cause of the disease and a need for new mechanistic or targeted techniques. A significant characteristic of HCC, in common with other aggressive cancers, is the presence of reprogrammed, hyperactive cell metabolism. Tumor cells hijack metabolic pathways to promote their uncontrolled growth, stress survival, invasion and metastasis. While classical mechanisms such as the Warburg effect, lipid metabolism and glutamine utilization have been understood, the lysosome, which was once viewed as a static “waste disposal unit” to remove old organelles and proteins, is instead a dynamic signaling and metabolic core. The lysosomes incorporate nutrients, energy and stress signals by master regulators such as mTORC1 (activated on its surface) that balance anabolic growth and catabolic recycling to the cellular demands. In HCC, lysosomes are not passive, but are highly active and dysregulated. HCC cells upregulate lysosomes, which scavenge intracellular components via enhanced autophagy and engulf extracellular proteins via macropinocytosis, crucial for survival in the nutrient-poor, hypoxic tumor microenvironment. In addition to metabolism, lysosomes exhibit pro-invasive functions by secreting hydrolases to remodel the extracellular matrix, promote angiogenesis, and suppress stromal immune cells to foster a pro-tumor microenvironment. In a clinical context, lysosomes play an important role in therapeutic resistance: they sequester and inactivate chemotherapeutics via lysosomal sequestration, and enhanced autophagic flux protects the cell from therapy-induced damage, contributing to relapse, as lysosomal dysfunction is a key cause of treatment failure. This makes lysosomes promising yet challenging therapeutic targets in HCC. Recent preclinical and early clinical studies investigate multiple strategies to exploit the susceptibility of lysosomes: lysosome-specific agents, alkalinizing the lysosome lumen or inducing membrane permeabilization and lysosome-dependent cell death; pharmacological inhibition of key lysosomal enzymes or autophagy to impair nutrient recycling and stress adaptation; smart nanotherapeutic agents or antibody-drug conjugates, specifically activated in the acidic lysosomal environment or utilizing lysosomal pathways for efficient intracellular drug release; and combination strategies of lysosome-targeting agents with tyrosine kinase inhibitors or immunotherapy to overcome resistance and achieve synergistic antitumor effects. In summary, our review systematically presents the role of lysosomes in HCC, from metabolic reprogramming and microenvironmental adaptation to therapeutic resistance. By synthesizing the latest mechanistic insights and preclinical advances, this review highlights the indispensable role of lysosomes in the complex HCC biological network, emphasizing that an in-depth understanding of this dynamic organelle holds great promise for developing innovative, targeted therapies, offering new hope for improving the poor prognosis of global HCC patients.
2.Staged Efficacy of Qijia Rougan Prescription Combined with Entecavir for Chronic Hepatitis B-related Hepatic Fibrosis with Qi Deficiency and Collateral Stasis Syndrome Based on "Zhu Ke Jiao" Theory
Baixue LI ; Xin WANG ; Jibin LIU ; Li WEN ; Cen JIANG ; Wenjun WU ; Dong WANG ; Shuwan LIU ; Huabao LIU ; Yongli ZHENG ; Liang HUANG ; Yue SU ; Song ZHANG ; Yanan SHANG ; Hang ZHOU ; Quansheng FENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):180-188
ObjectiveThis paper aims to investigate and evaluate the staged efficacy and safety of the representative empirical prescription of the “Zhu Ke Jiao” theory, Qijia Rougan prescription, combined with entecavir in the treatment of hepatic fibrosis in chronic hepatitis B. MethodsA multicenter randomized controlled clinical study was conducted, and 101 patients diagnosed with chronic hepatitis B-related hepatic fibrosis (CHB-HF) who met the diagnosis and inclusion criteria were randomly assigned to an observation group (Qijia Rougan prescription + entecavir) and a control group (entecavir). The treatment duration was 24 weeks. Liver stiffness measurement (LSM), fibrosis-4 index (FIB-4), portal vein diameter, hepatitis B serology, biochemical indicators, hepatic fibrosis markers in serum [hyaluronic acid (HA), laminin (LN), procollagen Ⅲ peptide (PⅢP), and type Ⅳ collagen (Ⅳ-C)], and traditional Chinese medicine syndrome scores were used as efficacy evaluation indicators. Efficacy assessments and explorations of different staged subgroups of Qijia Rougan prescription were conducted according to LSM values based on the Metavir pathological staging standard. ResultsA total of 98 cases were included for statistical analysis, with 49 cases in the observation group and 49 in the control group. The general data of the patients in both groups were comparable. Compared with the same group before treatment, the observation group showed a significant reduction in LSM and FIB-4 (P<0.01), as well as notable improvements in LN, Ⅳ-C, and various TCM syndrome scores (P<0.05, P<0.01). When compared to the control group after treatment, the observation group demonstrated significant improvements in LSM, FIB-4, and various TCM syndrome score indicators (P<0.05, P<0.01), indicating that the observation group performed better than the control group. Subgroup analysis of the regression of hepatic fibrosis stages showed that compared to the same group before treatment, the observation group had better improvement in regression of stages F2 and F3 (P<0.05). When compared to the control group after treatment, the observation group exhibited superior improvement in regression of stage F3 (P<0.05). No adverse events occurred in either group during the treatment period. ConclusionCompared with entecavir alone, the combination of Qijia Rougan prescription and entecavir significantly improves the degree of hepatic fibrosis and clinical TCM symptoms in patients. The optimal intervention period is primarily during stage F3, which is a potential “interception” point of the “Zhu Ke Jiao” theory.
3.Quantitative Molecular Detection of Angelicae Sinensis Radix and Its Processed Products Based on Herb-Q Method
Mingyu ZHANG ; Wenjun JIANG ; Baoyu JI ; Yue WANG ; Haitao ZHANG ; Haobo ZHANG ; Xue FENG ; Xiwen LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):192-200
ObjectiveAngelicae Sinensis Radix, a commonly used medicinal herb with both medicinal and edible properties, is frequently adulterated in the market, severely affecting the clinical efficacy of preparations. While qualitative identification techniques for adulterants and counterfeits are now relatively mature, quantitative detection methods for adulterated processed products remain unexplored. Quantitative detection research of Angelicae Sinensis Radix and its primary closely related adulterant, "Tu Danggui" (Angelica gigas), was conducted to establish a herbal quantitative molecular detection (Herb-Q) method for Angelicae Sinensis Radix and its processed products, providing a model for the establishment of quantitative detection technologies for Angelicae Sinensis Radix and related health products. MethodsThe specific single-nucleotide polymorphism (SNP) loci of Angelicae Sinensis Radix and Angelica gigas Nakai were screened based on the complete chloroplast genome sequence. The specific SNP loci of Angelicae Sinensis Radix were selected for quantitative methodological investigations (linearity, limit of quantification, limit of detection, and reproducibility) by mixing the powder of the herbs with different adulteration ratios. Huoxue Zhitong powder with three distinct adulteration ratios (15%, 25%, and 35%) was utilized to ascertain the precision of the Herb-Q method for the quantitative detection of Chinese patent medicines containing Angelicae Sinensis Radix. ResultsBy comparing the 123 chloroplast genome sequences of Angelicae Sinensis Radix, based on the principles of intraspecies conservation, interspecies specificity, and meeting the requirements of pyrophosphate high-throughput sequencing, it was determined that 9 674th locus (A/G) in the chloroplast genome sequence NC_042826.1 and 38 592nd locus (T/C) in the chloroplast genome sequence NC_029393.1 could be the exclusive molecular identification loci of Angelicae Sinensis Radix and Angelica gigas Nakai, respectively. The linear relationship R2 of the Herb-Q method established by selecting the specific 9 674th locus (A/G) of Angelicae Sinensis Radix was 0.997 4 (R2>0.99), indicating an excellent linear relationship. The limits of quantification and detection were established at 2.0%, exhibiting excellent reproducibility [relative standard deviation(RSD)<2.0%]. The established quantitative system based on the Herb-Q method detected the adulteration amount of counterfeit A. gigas in the Huoxue Zhitong powder, with an average deviation of 1.3% for three molecular quantitative replicates. ConclusionThis research demonstrates that the Herb-Q quantitative detection method established based on the 9 674th locus (A/G) in the chloroplast genome sequence NC_042826.1 of Angelicae Sinensis Radix has good applicability, objectivity, and accuracy for Angelicae Sinensis Radix and A. gigas, and its processed products. This method has the capacity to provide technical support for the quantitative detection of commercially available Angelicae Sinensis Radix derivatives, including traditional Chinese medicinal preparations, dietary supplements, and nutraceuticals.
4.Quantitative Molecular Detection of Angelicae Sinensis Radix and Its Processed Products Based on Herb-Q Method
Mingyu ZHANG ; Wenjun JIANG ; Baoyu JI ; Yue WANG ; Haitao ZHANG ; Haobo ZHANG ; Xue FENG ; Xiwen LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):192-200
ObjectiveAngelicae Sinensis Radix, a commonly used medicinal herb with both medicinal and edible properties, is frequently adulterated in the market, severely affecting the clinical efficacy of preparations. While qualitative identification techniques for adulterants and counterfeits are now relatively mature, quantitative detection methods for adulterated processed products remain unexplored. Quantitative detection research of Angelicae Sinensis Radix and its primary closely related adulterant, "Tu Danggui" (Angelica gigas), was conducted to establish a herbal quantitative molecular detection (Herb-Q) method for Angelicae Sinensis Radix and its processed products, providing a model for the establishment of quantitative detection technologies for Angelicae Sinensis Radix and related health products. MethodsThe specific single-nucleotide polymorphism (SNP) loci of Angelicae Sinensis Radix and Angelica gigas Nakai were screened based on the complete chloroplast genome sequence. The specific SNP loci of Angelicae Sinensis Radix were selected for quantitative methodological investigations (linearity, limit of quantification, limit of detection, and reproducibility) by mixing the powder of the herbs with different adulteration ratios. Huoxue Zhitong powder with three distinct adulteration ratios (15%, 25%, and 35%) was utilized to ascertain the precision of the Herb-Q method for the quantitative detection of Chinese patent medicines containing Angelicae Sinensis Radix. ResultsBy comparing the 123 chloroplast genome sequences of Angelicae Sinensis Radix, based on the principles of intraspecies conservation, interspecies specificity, and meeting the requirements of pyrophosphate high-throughput sequencing, it was determined that 9 674th locus (A/G) in the chloroplast genome sequence NC_042826.1 and 38 592nd locus (T/C) in the chloroplast genome sequence NC_029393.1 could be the exclusive molecular identification loci of Angelicae Sinensis Radix and Angelica gigas Nakai, respectively. The linear relationship R2 of the Herb-Q method established by selecting the specific 9 674th locus (A/G) of Angelicae Sinensis Radix was 0.997 4 (R2>0.99), indicating an excellent linear relationship. The limits of quantification and detection were established at 2.0%, exhibiting excellent reproducibility [relative standard deviation(RSD)<2.0%]. The established quantitative system based on the Herb-Q method detected the adulteration amount of counterfeit A. gigas in the Huoxue Zhitong powder, with an average deviation of 1.3% for three molecular quantitative replicates. ConclusionThis research demonstrates that the Herb-Q quantitative detection method established based on the 9 674th locus (A/G) in the chloroplast genome sequence NC_042826.1 of Angelicae Sinensis Radix has good applicability, objectivity, and accuracy for Angelicae Sinensis Radix and A. gigas, and its processed products. This method has the capacity to provide technical support for the quantitative detection of commercially available Angelicae Sinensis Radix derivatives, including traditional Chinese medicinal preparations, dietary supplements, and nutraceuticals.
5.Analysis of the association between chronic liver disease and depressive disorder and the mediating effect of nocturnal sleep duration
Panshili HAN ; Yue FENG ; Yanhang GAO
Journal of Clinical Hepatology 2026;42(4):890-899
ObjectiveTo investigate the association between chronic liver disease (CLD) and depression and the mediating role of nocturnal sleep duration, as well as the consistency of this association between Chinese and American populations. MethodsThe data from two databases were integrated, i.e., China Health and Retirement Longitudinal Study (CHARLS) (n=14 225) in China and National Health and Nutrition Examination Survey (NHANES) (n=11 353) in the United States, and the subjects were divided into CLD group and non-CLD group. A stepwise Logistic regression model was used to assess the independent association between CLD and depression. Bootstrap resampling (1 000 times) was used to quantify the proportion of the mediating effect of nocturnal sleep duration, and interaction effects were evaluated through stratified analyses based on the factors including registered residence/race and income. The independent-samples t test was used for comparison of continuous data between two groups; the chi-square test or the Fisher’s exact test was used for comparison of categorical data; a multivariate Logistic regression model analysis was used to investigate the association of different types of CLD with depression. ResultsAfter multivariate adjustment, CLD was significantly associated with the increased risk of depression (CHARLS: odds ratio [OR]=1.76, 95% confidence interval [CI]: 1.46 — 2.12, P<0.001; NHANES: OR=1.87, 95%CI: 1.50 — 2.35, P<0.001). Nocturnal sleep duration played a partial mediating role in the association between CLD and depression, with a mediating effect accounted for 12.41% in CHARLS and 2.16% in NHANES (P<0.05). The interaction analysis showed that in CHARLS, the association between CLD and depression was more significant in the residents with agricultural registered permanent residence (OR=2.07, 95%CI: 1.67 — 2.57, P<0.001), and in NHANES, this association was more significant in the population with moderate poverty income ratio (OR=2.66, 95% CI: 1.92 — 3.69, P<0.001). The subtype analysis showed that autoimmune hepatitis (OR=3.63, 95%CI: 1.49 — 8.88, P=0.005), liver cirrhosis (OR=2.46, 95%CI: 1.32 — 4.60, P=0.005), and fatty liver disease (OR=1.75, 95%CI: 1.27 — 2.39, P=0.001) significantly increased the risk of depression, while no statistical significance was observed for viral hepatitis or other liver diseases (P>0.05). ConclusionThis study reveals the significant association between CLD and depression at the population level and suggests that nocturnal sleep duration may play a partial mediating role, which is consistent between the Chinese and American populations. These research findings provide quantitative evidence-based support for understanding the underlying mechanism of CLD-related depression and points out the potential significance of sleep issues in CLD management.
6.Construction and Evaluation of Animal Model with "Phlegm-dampness" Syndrome
Xiaoqin LIU ; Qingzhi LIANG ; Wei JIANG ; Ling DENG ; Haoyue FENG ; Rensong YUE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):26-39
According to traditional Chinese medicine (TCM) theory, impaired spleen transportation function disrupts nutrient distribution, causing metabolic accumulation of lipids that transform into pathogenic phlegm-dampness. These pathological factors disseminate through the San Jiao and obstruct meridian pathways, ultimately forming the pathogenesis described as "all disorders involve phlegm". Phlegm and dampness share common pathogenic origins but manifest distinct clinical manifestations. Dampness, as the precursor, may congeal into phlegm, while existing phlegm accumulation can further exacerbate dampness stagnation, thereby establishing a self-perpetuating pathological cycle. Modern medical research has confirmed that the essence of "phlegm-dampness" syndrome is closely associated with energy metabolism disorders, serving as a common pathological basis for metabolic syndrome, type 2 diabetes mellitus, atherosclerosis, and other major chronic diseases. As a crucial vehicle for medical experimental research, disease-syndrome combination animal models serve as an indispensable means to advance the modernization of TCM. Currently, based on classical theories such as "rich and greasy foods produce phlegm" and "physical coldness combined with cold consumption causes external pathogens to invade the skin and hair, thereby generating internal dampness", researchers primarily employ two paradigms to construct animal models of phlegm-turbidity, dampness obstruction, and phlegm-dampness syndromes: the first involves simulating TCM etiological factors (through methods like dietary irregularities, imblanace between work and rest, and combined internal-external dampness exposure), while the second combines disease with syndrome differentiation (inducing pathological changes through physical, chemical, or biological interventions). Through comprehensive evaluation incorporating macroscopic observation and microscopic index detection, model animals undergo systematic biological and pathological assessment, with further syndrome type verification achieved via the "prescription-based syndrome detection" approach. However, existing models still exhibit significant deficiencies in both the standardization of modeling methodologies and the systematization of evaluation criteria. This paper reviews the strategies for constructing "phlegm-dampness" syndrome animal models and their corresponding evaluation indices, focusing on the pathological correlations among different modeling approaches. The aim is to provide methodological guidance for research on TCM syndromes related to "phlegm-dampness" syndrome and to support the development of TCM therapies for resolving phlegm and eliminating dampness. This study not only contributes to advancing the standardization of TCM syndrome research but also provides crucial technical support for the modernization of TCM.
7.DIA Proteomic Profiling on Staged Regulatory Effect of Tonifying Deficiency and Dredging Collaterals Method on Liver Fibrosis in Rats Based on Theory of "Zhu Ke Jiao"
Xin WANG ; Pengyu ZHU ; Li WEN ; Jibin LIU ; Aochun YUE ; Ziyi CHEN ; Jing ZHANG ; Li ZHU ; Quansheng FENG ; Cen JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):119-132
ObjectiveThis paper aims to investigate the differential mechanisms underlying the staged therapeutic effects of Qijia Rougan formula on liver fibrosis using proteomic technology. MethodsThe staged rat model of liver fibrosis was established by subcutaneous injection of carbon tetrachloride (CCl4) and olive oil. One hundred and four SD rats were randomized into thirteen groups:a normal group,a two-week model group,a four-week model group,a six-week model group,an eight-week model group,a two-week Qijia Rougan formula group,a four-week Qijia Rougan formula group,a six-week Qijia Rougan formula group,an eight-week Qijia Rougan formula group,a two-week compound Biejia Ruangan tablet group,a four-week Compound Biejia Ruangan Tablet group,a six-week Compound Biejia Ruangan Tablet group,and an eight-week compound Biejia Ruangan tablet group. After two weeks of drug intervention,liver tissue and abdominal aortic blood samples were collected from the rats for testing. Hematoxylin-eosin (HE) staining,Masson staining,and Picro Sirius red staining were used to observe pathological damage and collagen fiber deposition in liver tissues. Immunohistochemistry (IHC) was employed to detect the contents of fibrosis markers in liver tissues. The contents of liver function indicators in the serum were measured using a fully automated biochemical analyzer,and the levels of liver fibrosis indicators in the serum were assessed by enzyme-linked immunosorbent assay (ELISA). Liver tissues from the normal group,each model group,and each Qijia Rougan formula group were subjected to label-free quantitative proteomic analysis to identify differential proteins among the groups,with key proteins validated by Western blot. Finally,bioinformatics analysis was performed on the differential proteins. Results(1) The staged rat model of liver fibrosis constructed with CCl4 and olive oil showed pathological results at the 2nd,4th,6th,and 8th weeks of modeling that were consistent with the Metavir standards for the F1,F2,F3,and F4 stages. Compared with those in the normal control group,the protein expressions of α-smooth muscle actin (α-SMA) and Collagen Ⅰ were significantly increased in each stage (P<0.05). The levels of liver function indicators in the serum,including alanine aminotransferase (ALT),aspartate aminotransferase (AST),alkaline phosphatase (ALP),direct bilirubin (DBIL),and total bilirubin (TBil) in each model group,were significantly elevated in each stage (P<0.01). The levels of liver fibrosis indicators in the serum,including procollagen Ⅲ peptide (PⅢP),type Ⅳ collagen(Ⅳ-C),hyaluronic acid (HA),and laminin (LN) in each model group,were significantly increased in each stage (P<0.05,P<0.01). This study successfully established a staged rat model of liver fibrosis. (2) Compared with the model groups at each stage,the administration groups showed a reduction in hepatocyte ballooning degeneration,a more orderly arrangement of hepatocytes,and a decrease of inflammatory cell infiltration. The blue-stained collagen fibers became significantly thinner and finer,with reduced and narrowed fibrous septa. The areas of collagen fibers and Picro Sirius red staining were reduced (P<0.05). The positive areas of α-SMA and Collagen Ⅰ expression were significantly decreased (P<0.05). The levels of ALT,AST,ALP,DBIL,and TBil in the rats of the model groups at each stage were significantly reduced (P<0.05,P<0.01). The levels of PⅢP,Ⅳ-C,HA,and LN in the rats of the model groups at each stage were significantly decreased (P<0.05). Among these,the improvements in all indicators were most significant in the F3 stage (P<0.01).(3) The proteomic results show that a total of 165 differential proteins exhibit a callback trend when comparing the model groups at four stages with the normal group,and when comparing the Qijia Rougan formula group with the model group. Western blot analysis reveals that the levels of NAD(P)H:quinone oxidoreductase 1 (NQO1),mitogen-activated protein kinase 1 (MAPK1),arginase 1 (Arg1),and glutathione S-transferase α1 (GSTA1) were consistent with the proteomic results. Bioinformatics results reveal that 165 differentially expressed proteins are enriched in multiple signaling pathways. Notably,signaling pathways such as drug metabolism-cytochrome P450,arginine biosynthesis,and the peroxisome proliferator-activated receptor (PPAR) signaling pathway were found to be closely associated with liver fibrosis,suggesting that the Qijia Rougan formula may exert its staged regulatory effects on liver fibrosis by regulating these pathways. ConclusionThe Qijia Rougan formula may achieve staged regulation of liver fibrosis by regulating drug metabolism-cytochrome P450,arginine biosynthesis,and the PPAR signaling pathway.
8.The Role of Circulating Tumor Cell as a Promising Biomarker in the Evaluation of Pulmonary Nodules: A Prospective Study
Shijie WANG ; Changdan XU ; Xiaohong XU ; Weipeng SHAO ; Guohui WANG ; Xiongtao YANG ; Liwei GAO ; Feng TENG ; Hongliang SUN ; Yue ZHAO ; Hongxiang FENG ; Guangying ZHU
Cancer Research and Treatment 2026;58(1):128-140
Purpose:
Our previous study showed that circulating tumor cell (CTC) count combined with gene mutation detection might help differentiate benign and malignant pulmonary nodules (PNs). Herein, we aimed to expand the study cohort and conduct further sequencing analysis.
Materials and Methods:
Patients with PNs were included, and CTCs were identified before operation. Low-coverage whole-genome sequencing (LC-WGS) and lung cancer-related targeted gene sequencing were performed on CTCs. The diagnostic efficacy was evaluated by receiver operating characteristic (ROC) curve. The differences in CTC counts among subgroups classified by demographic–clinical characteristics were analyzed. LC-WGS–based copy number variation (CNV) analysis and targeted gene mutation analysis were conducted.
Results:
A total of 172 patients were included. CTC count of 2.5 was identified by the ROC curves as the optimal diagnostic cutoff. The sensitivity and specificity of CTC count for differentiating benign and malignant PNs were 54.2% and 78.6%, respectively. The diagnostic sensitivity and specificity of combined CTC count, radiological nodule type, and any malignant imaging features were 84.7% and 71.4%, respectively. The CTC counts were significantly greater in patients with aggressive tumors, later stage, and spread through air spaces. CTCs from malignant cases had more CNVs than those from benign cases.
Conclusion
CTC count can be used in identifying malignant PNs. The diagnostic efficacy can be improved if combined with computed tomography imaging characteristics. Further CNV analysis might help differential diagnosis. Greater CTC count might suggest more aggressive tumors. CTC detection can provide important information and guidance for subsequent management of PNs.
9.Applications and advancements of artificial intelligence in removable partial dentures
FENG Yue ; WANG Fu ; FENG Zhihong ; NIU Lina
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(7):631-641
With the rapid aging of the global population, the demand for prosthodontic rehabilitation of partial edentulism continues to increase. Among the available treatment options, removable partial dentures remain an essential clinical modality for restoring partially edentulous arches and masticatory function because of their broad indications, minimal invasiveness, and cost-effectiveness. However, unlike fixed prosthodontics, which have undergone substantial digitalization, removable partial denture therapy is characterized by dual tooth-tissue support, complex biomechanics, and highly variable anatomical morphology. As a result, the digital design and fabrication of removable partial dentures have long faced major bottlenecks, including limited standardization, steep learning curves, and heavy dependence on expert experience. In recent years, artificial intelligence (AI) has undergone a paradigm shift from early expert systems to machine learning and deep learning, creating new opportunities to address the complexity and precision demands of removable partial denture design. Based on a comprehensive literature review and our team's clinical experience, this article systematically elaborates on the specific applications and normative standards of AI throughout the entire removable partial denture fabrication workflow. In the data acquisition phase, we emphasize multimodal data fusion and digital pressure impression strategies. In the intelligent diagnosis and planning phase, deep learning is utilized to achieve the automatic identification of edentulous areas and undercuts, while machine learning and expert systems assist in abutment selection and treatment plan generation. Regarding automated design logic, the article analyzes the process from the intelligent planning of the common path of insertion to parametric design based on biomechanical optimization. At the clinical operation level, a standardized human-machine collaborative workflow of “AI generation-clinician review-local fine-tuning” is proposed. In terms of ethics and accountability, the principles of algorithmic transparency and “the clinician as the ultimate responsible subject” are emphasized. Furthermore, integrating clinical practice, this article innovatively proposes a “capability grading system for AI-assisted removable partial denture design (comprising the auxiliary analysis level, the rule-based design level, and the intelligent adaptive level)”. The aim is to offer practical recommendations for dental clinicians, dental technicians, and researchers, thereby facilitating the transition of removable partial denture prosthodontics toward greater precision, intelligence, and standardization.
10.Cloning and Functional Characterization of Farnesyl Diphosphate Synthase Gene in Biosynthesis of Terpenoid Components in Chinese Materia Medica
Yue ZHANG ; Feng ZHANG ; Yue ZHANG ; Chaoyue LIU ; Bolin ZHANG ; Jia LIU ; Caixia WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):175-183
ObjectiveThis study aims to enhance of the farnesyl pyrophosphate(FPP) pool in Saccharomyces cerevisiae by heterologously expressing different farnesyl diphosphate synthases(FPSs) from various plants, thereby increasing the production of terpenoid compounds by the engineered yeast. MethodsRNA from mixed samples of roots, stems, and leaves of seven plants including Arabidopsis thaliana, Rosa rugosa, Artemisia annua, Centella asiatica, Humulus lupulus, Medicago sativa, and Panax ginseng was extracted by column chromatography and reverse transcribed into the first strand of complementary DNA(cDNA), and based on the transcriptome data of the seven species of plants, sequence-specific primers were designed for CaFPS, RrFPS, MsFPS, HiFPS, PgFPS, AtFPS, and AaFPS, the full-length of the genes was cloned, and the genes were analyzed for bioinformatics in order to construct a pESC yeast shuttle vector. These seven plant-derived FPSs were further heterologously expressed in the previous constructed β-elemene-producing yeast, and the yield of β-elemene was indicated for their catalytic acivities. ResultsThe coding sequences of CaFPS, RrFPS, MsFPS, HiFPS, PgFPS, AtFPS, and AaFPS were all of 1 021 bp in length and encoding 301 amino acids, all of which were similarly related to the endogenous FPS-encoding gene(ERG20) in S. cerevisiae. After heterologous expression, RrFPS was identified as the most effective in catalyzing the synthesis of FPP from isopentenyl pyrophosphate(IPP) and dimethylallyl pyrophosphate(DMAPP). Compared to the control strains, the RrFPS overexpressed yeast strains YB-1-Rr and YB-3-Rr increased the production of β-elemene by 231.25% and 189.3%, respectively. ConclusionBy comparing the functions of FPS-encoding genes from seven different plant sources, it is determined that the protein encoded by the RrFPS from R. rugosa has the best catalytic ability, which can provide key genetic elements for the construction of engineered yeast strain constructs with high terpenoid production.


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