1.Prognostic analysis of genes related to pyroptosis in prostate cancer cells and the regulatory role of NLRP1
Xiaolu MA ; Jiaqin CHEN ; Junlong FENG ; Qi ZHAO ; Bin WANG
Journal of Modern Urology 2025;30(1):73-81
[Objective] To analyze the prognostic value of prostate cancer (PCa) pyroptosis-related genes (PRGs) using gene expression databases and to explore the regulatory mechanism of nucleotidebinding oligomerization domain-like receptor containing pyrin domain 1 (NLRP1) in the pyroptosis of PCa cells. [Methods] Fragments per kilobase of exon model per million reads mapped (FPKM) data and clinical information from PCa and adjacent tissues from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were obtained. Differentially expressed PRGs between PCa and adjacent tissues, classified subtypes and plotted survival curves were analyzed. Univariate Cox regression analysis, least absolute shrinkage and selection operator (LASSO) regression analysis were conducted to screen prognosis-related PRGs, risk scores were calculated, and a prognostic risk model was constructed and validated. Patients were divided into high and low risk groups based on the median risk scores from the training and validation sets, and gene ontology (GO) enrichment and kyoto encyclopedia of genes and genomes (KEGG) analysis were conducted on differentially expressed PRGs. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression level of NLRP1 in PCa cell lines, and pyroptosis was induced in DU145 and LNCaP cells while morphological changes were observed. Western blot (WB) was performed to detect the expression of pyroptosis-related molecules. [Results] A total of 6 prognostic-related PRGs were obtained, including CHMP4C, CYCS, GPX4, GSDMB, NLRP1, and PLCG1. The risk score was positively correlated with the risk of recurrence but negatively correlated with the progression-free survival (P<0.001). The area under the receiver operating characteristic curves (AUCs) for the training set at 1, 3, and 5 years were 0.769 (95%CI: 0.652-0.878), 0.804 (95%CI: 0.736-0.882), and 0.772 (95%CI: 0.631-0.905), respectively, while those for the validation set were 0.731 (95%CI: 0.647-0.826), 0.753 (95%CI: 0.674-0.818), and 0.763 (95%CI: 0.626-0.849), respectively. Differences in expression levels of the 6 PRGs were observed between the high and low risk groups in both the training and validation sets (P<0.05). Cox regression analysis showed that T stage, prostate specific antigen (PSA), Gleason grade, and risk score were independent predictors of PCa prognosis (P<0.05). Differences in risk scores were observed among patients of different ages, T stages, and Gleason grades (P<0.05). NLRP1 was found to be lowly expressed in PCa cell lines and was involved in the regulation of pyroptosis in DU145 and LNCaP cells. [Conclusion] The prognostic risk model constructed based on PRGs has a certain predictability for the prognosis of PCa patients, and NLRP1 may be involved in the regulation of pyroptosis in PCa cells.
2.Mechanism of Gushining Granules in Attenuating Dexamethasone-induced Apoptosis of Bone Marrow Mesenchymal Stem Cells via Activating PI3K/Akt/Bad Signalling Pathway
Chengyu CHU ; Lei ZHU ; Long LIANG ; Feng WANG ; Xuejian YU ; Wenwu LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):115-122
ObjectiveTo establish steroid-induced osteonecrosis of the femoral head (SANFH) cell model by using dexamethasone (DEX)-induced bone marrow mesenchymal stem cells (BMSCs) and demonstrate that Gushing Granules (GSNs) exert an improving effect by activating the phosphatidylinositol-3-kinase/protein kinase B/B-lymphoma-2 gene related promoter (PI3K/Akt/Bad) signalling pathway. MethodsFirstly, SD rats were orally administered with drugs at a dose of 0.9 g·kg-1 to prepare GSN-containing serum, and CCK-8 screening was used to determine the optimal dosage and duration of action. Then, BMSCs were cultured and treated with 1×10-6 mol·L-1 DEX, 10% GSN-containing serum, and inhibitor LY294002 of PI3K/Akt signalling pathway for 24 hours to model and group SANFH cells. Cell viability and proliferation were detected by using CCK-8 assay kit and EdU staining kit. Flow cytometry was used to detect cell apoptosis. An alkaline phosphatase (ALP) assay kit was employed to detect ALP expression. In order to detect the PI3K/Akt/Bad signalling pathway and protein and mRNA expression of apoptosis-related proteins such as apoptosis regulatory factors B-cell lymphoma-2 gene (Bcl-2), and Bcl-2-associated X protein (Bax), osteocalcin (OCN), and Collagen Ⅰ, we used Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultsThe CCK-8 assay kit determined that the optimal dosage for GSN-containing serum is 10%, and the duration of action is 48 hours. After modelling and grouping the cells in each group, the detection results showed that the SANFH model group had significantly lower cell viability, cell proliferation, and ALP expression, as well as protein and mRNA expressions of PI3K, Akt, Bad, Bcl-2, OCN, and Collagen I compared to the blank group. The nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry significantly increased (P<0.05,P<0.01). After treatment with GSN-containing serum, cell viability, cell proliferation, and ALP expression, as well as expressions of PI3K, Akt, Bad, Bcl-2, OCN, and Collagen Ⅰ nucleic acids and proteins were significantly increased, while the nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry significantly decreased(P<0.05,P<0.01). Compared with the GSN drug-containing serum group, the simultaneous treatment with the inhibitor LY294002 and GSN drug-containing serum reversed the improvement effect of GSN. Specifically, the cell viability, cell proliferation, ALP expression, and the nucleic acid and protein levels of PI3K, Akt, Bad, Bcl-2, OCN, and Collagen Ⅰ were all significantly decreased, while the nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry were significantly increased (P<0.05, P<0.01). ConclusionGSNs antagonize DEX-induced apoptosis of BMSCs by activating the PI3K/Akt/Bad signalling pathway, providing a scientific theoretical basis for the clinical treatment of SANFH with GSNs.
3.Molecular epidemiological investigations of human parvovirus B19 among blood donors in Lanzhou
Kangle WU ; Shulong YANG ; Weiping FENG ; Weirong WANG ; Deng PAN
Chinese Journal of Blood Transfusion 2025;38(1):68-72
[Objective] To investigate the molecular prevalence and genotype of human parvovirus B19(B19) among blood donors in Lanzhou, and provide data support for monitoring the positive rate of B19 DNA in local blood donors. [Methods] A total of 7 644 blood donor samples collected from January to September 2022 were randomly screened using real-time fluorescent PCR, resulting in 23 samples testing positive for B19 DNA. The characteristics of the B19 DNA reactive donors including gender, age, blood donation recruitment and promotion mode, and donation frequency were analyzed using SPSS 22.0. Additionally, the VP1 gene fragment of B19 DNA reactive samples was sequenced and an evolutionary tree was constructed by the N-J method. [Results] The results showed that the positive rate of B19 DNA in Lanzhou was 0.30%, and the positive population mainly consisted of female individuals aged 18-30 years old who were first-time blood donors; furthermore, genotype 1a was identified as predominant. [Conclusion] The positive rate of B19 DNA is low among blood donors in Lanzhou, with genotype 1a being predominant. It is recommended to periodically monitor the B19 prevalence in blood donors and enhance prevention and control measures, thus improving blood quality and safety.
4.Optimization of drug management model for investigator-initiated trial with benchmarking analysis
Yufei XI ; Tianxiao WANG ; Xue ZHANG ; Yingzhuo DING ; Li YAN ; Feng JIANG ; Xiangui HE ; Jiannan HUANG ; Qin LI
China Pharmacy 2025;36(3):280-284
OBJECTIVE To optimize the management model of drugs used in investigator-initiated trial (IIT). METHODS With benchmarking analysis, based on the practical work experience of a tertiary specialized hospital in the field of IIT drug management in Shanghai, a thorough review was conducted, involving relevant laws, regulations, and academic literature to establish benchmark criteria and the evaluation standards. Starting from the initiation of IIT projects, a detailed comparative analysis of key processes was carried out, such as the receipt, storage, distribution, use and recycling of drugs for trial. The deficiencies in the current management of IIT drugs were reviewed in detail and a series of optimization suggestions were put forward. RESULTS It was found that the authorized records of drug management were missing, the training before project implementation was insufficient, and the records of receipt and acceptance of IIT drugs were incomplete. In light of these existing problems, improvement measures were put forward, including strengthening the training of drug administrators and stipulating that only drug administrators with pharmacist qualifications be eligible to inspect and accept drugs, etc. The related systems were improved, and 17 key points of quality control for the management of IIT drugs were developed. CONCLUSIONS A preliminary IIT drug management system for medical institutions has been established, which helps to improve the institutional X2023076) framework of medical institutions in this field.
5.Therapeutic Effect and Mechanism of Shentong Zhuyutang Combined with Dilongtang in Treatment of Lumbar Disc Herniation with Qi Stagnation and Blood Stasis Syndrome
Huangsheng TAN ; Yinbo WANG ; Yong HUANG ; Juyi LAI ; Hualong FENG ; Zhiming LAN ; Yuanfei FU ; Yong JIANG ; Shenghua HE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):47-54
ObjectiveTo observe the clinical efficacy of Shentong Zhuyutang combined with Dilongtang in the treatment of lumbar disc herniation (LDH) with Qi stagnation and blood stasis syndrome, and its effect on nucleus pulposus reabsorption and immune-inflammatory factors, exploring its therapeutic mechanism from the perspective of reabsorption. MethodsA total of 120 patients with LDH from the Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, treated between June 2020 and January 2023, were randomly divided into the control group (52 cases, with 8 dropouts) and the observation group (49 cases, with 11 dropouts) according to a random number table. The control group received routine treatment, while the observation group was treated with Shentong Zhuyutang combined with Dilongtang in addition to routine treatment. Visual Analogue Scale (VAS), Oswestry Disability Index (ODI), Japanese Orthopaedic Association (JOA) score, and traditional Chinese medicine (TCM) syndrome score were measured before treatment and after 3 courses of treatment. Venous blood samples were collected for the determination of serological indexes. MR examination was performed during the 6-month follow-up to calculate the absorption rate. ResultsAfter treatment, both groups showed significant reductions in VAS, ODI, TCM syndrome score, serum tumor necrosis factor (TNF)-α, matrix metalloproteinase (MMP)-9, and vascular endothelial growth factor (VEGF) levels, and a significant increase in JOA score compared with pre-treatment values (P<0.05). Compared with the control group, the observation group showed significantly lower VAS, ODI, TCM syndrome score, serum TNF-α, MMP-9, and VEGF levels, and a significantly higher JOA score (P<0.05). The proportion of nucleus pulposus reabsorption in the observation group was 57.14% (28/49), significantly higher than 21.15% (11/52) in the control group (χ2=6.161, P<0.05). ConclusionShentong Zhuyutang combined with Dilongtang can effectively relieve pain, improve lumbar function, and alleviate TCM clinical symptoms in LDH patients with Qi stagnation and blood stasis syndrome. Imaging findings suggest that the treatment promotes the reabsorption of nucleus pulposus protrusion, while laboratory testing shows reduced serum levels of TNF-α, MMP-9, and VEGF, which contribute to the rehabilitation of patients.
6.Optimization Strategy and Practice of Traditional Chinese Medicine Compound and Its Component Compatibility
Zhihao WANG ; Wenjing ZHOU ; Chenghao FEI ; Yunlu LIU ; Yijing ZHANG ; Yue ZHAO ; Lan WANG ; Liang FENG ; Zhiyong LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):299-310
Prescription optimization is a crucial aspect in the study of traditional Chinese medicine (TCM) compounds. In recent years, the introduction of mathematical methods, data mining techniques, and artificial neural networks has provided new tools for elucidating the compatibility rules of TCM compounds. The study of TCM compounds involves numerous variables, including the proportions of different herbs, the specific extraction parts of each ingredient, and the interactions among multiple components. These factors together create a complex nonlinear dose-effect relationship. In this context, it is essential to identify methods that suit the characteristics of TCM compounds and can leverage their advantages for effective application in new drug development. This paper provided a comprehensive review of the cutting-edge optimization experimental design methods applied in recent studies of TCM compound compatibilities. The key technical issues, such as the optimization of source material selection, dosage optimization of compatible herbs, and multi-objective optimization indicators, were discussed. Furthermore, the evaluation methods for component effects were summarized during the optimization process, so as to provide scientific and practical foundations for innovative research in TCM and the development of new drugs based on TCM compounds.
7.Immunity-inflammation Mechanism of Viral Pneumonia and Traditional Chinese Medicine Treatment Based on Theory of Healthy Qi and Pathogenic Qi
Zheyu LUAN ; Hanxiao WANG ; Xin PENG ; Yihao ZHANG ; Yunhui LI ; Jihong FENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):239-247
Viral pneumonia is an infectious disease caused by virus invading the lung parenchyma and interstitial tissue and causing lung inflammation, with the incidence rising year by year. Traditional Chinese medicine (TCM) can treat viral pneumonia in a multi-component, multi-target, and holistic manner by targeting the core pathogenesis of pneumonia caused by different respiratory viruses, demonstrating minimal side effects and significant advantages. According to the theory of healthy Qi and pathogenic Qi in TCM, the struggle between healthy Qi and pathogenic Qi and the imbalance between immunity and inflammation run through the entire process of viral pneumonia, and the immunity-inflammation status at different stages of the disease reflects different relationships between healthy Qi and pathogenic Qi. Immune dysfunction leads to the deficiency of healthy Qi, causing viral infections. The struggle between healthy Qi and pathogenic Qi causes immunity-inflammation imbalance, leading to the onset of viral pneumonia. Inflammatory damage causes persistent accumulation of phlegm and stasis, leading to the progression of viral pneumonia. The cytokine storm causes immunodepletion, leading to the excess of pathogenic Qi and diminution of healthy Qi and the deterioration of viral pneumonia. After the recovery from viral pneumonia, there is a long-term imbalance between immunity and micro-inflammation, which results in healthy Qi deficiency and pathogenic Qi lingering. Healthy Qi deficiency and pathogenic Qi excess act as common core causes of pneumonia caused by different respiratory viruses. Clinical treatment should emphasize both replenishing healthy Qi and eliminating pathogenic Qi, helping to restore the balance between healthy Qi and pathogenic Qi as well as between immunity and inflammation, thus promoting the recovery of patients from viral pneumonia. According to the TCM theory of healthy Qi and pathogenic Qi, this article summarizes the immunity-inflammation mechanisms at different stages of viral pneumonia, and explores the application of the method of replenishing healthy Qi and eliminating pathogenic Qi in viral pneumonia. The aim is to probe into the scientific connotation of the TCM theory of healthy Qi and pathogenic Qi in viral pneumonia and provide ideas for the clinical application of the method of replenishing healthy Qi and eliminating pathogenic Qi to assist in the treatment of viral pneumonia.
8.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.
9.Analysis of Application of Animal Model of Spleen Deficiency and Dampness Syndrome Based on Data Mining
Qingqian YU ; Yifei ZHANG ; Zehan ZHANG ; Weiyue ZHANG ; Yuebo WANG ; Fengzhi WU ; Feng LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):235-243
ObjectiveThe research focuses on developing modeling and evaluation methodologies for an animal model exhibiting spleen deficiency and dampness excess syndrome, with the aim of standardizing such animal models for future reference. MethodsBy conducting a literature search on animal models of spleen deficiency and dampness excess syndrome, relevant publications meeting inclusion and exclusion criteria will be identified based on publication date, data source, types of diseases involved, animal characteristics, modeling methods, modeling duration, macroscopic syndrome assessment indicators, macroscopic quantification indicators, laboratory testing parameters, intervention approaches, positive controls and application context. A database will be established to facilitate the extraction of this information for quantitative analysis, statistical evaluation, and visual representation. ResultsA total of 137 literature articles meeting the standards have been included in the research. The primary animal species used in animal models of spleen deficiency and dampness excess are SD rats. Modeling methods include single-factor, dual-factor composite, and triple-factor composite methods, with various models widely applied in validation of pharmacological effects and mechanistic explorations. Evaluation indices of animal models for spleen deficiency and dampness excess primarily consist of macroscopic syndrome evaluation indicators and macroscopic quantitative indicators. Laboratory testing indicators are mostly related to research areas such as fluid metabolism and gastrointestinal function. The most commonly studied herbal formulas currently include Shenling Baizhu San and Pingwei San, with natural recovery and the use of the western medicine metronidazole as the most frequently used positive controls. ConclusionThe application of animal models for spleen deficiency and dampness excess is gradually increasing, with various modeling methods already simulating the typical characteristics of this syndrome pattern. However, there are still many areas that are worth contemplating and improving. This study aims to provide reference and ideas for the standardization of symptom names in animal models of spleen deficiency and dampness excess, as well as for the improvement of model construction and evaluation systems.
10.Trends in incidence and mortality of prostate cancer in Ningbo City from 2011 to 2023
ZHANG Qun ; WANG Yong ; CHEN Jieping ; BAO Kaifang ; FENG Yueyi ; Wang Xiaoli
Journal of Preventive Medicine 2025;37(1):46-50
Objective:
To analyze the trends in incidence and mortality of prostate cancer in Ningbo City, Zhejiang Province, so as to provide insights into the prevention and control of prostate cancer.
Methods:
Data of the incidence and mortality of prostate cancer in Ningbo City from 2011 to 2023 were collected through Ningbo Chronic Disease Collaborative Management System. The incidence and mortality of prostate cancer were calculated and standardized by the data from the sixth national population census in 2010 and the Segi's world standard population in 1960. The trends in incidence and mortality of prostate cancer were evaluated using average annual percent change (AAPC).
Results:
A total of 15 411 cases of prostate cancer were reported in Ningbo City from 2011 to 2023, and the crude incidence, Chinese-standardized incidence and world-standardized incidence were 39.62/105, 22.18/105 and 16.49/105, respectively, showing upward trends (AAPC=14.782%, 10.390% and 10.608%, all P<0.05). The Chinese-standardized incidence of prostate cancer was higher in urban areas than in rural areas, and both showed upward trends (25.14/105 vs. 19.44/105; AAPC=9.057% and 14.272%, both P<0.05). The crude incidence of prostate cancer in the groups aged 50-<60 years, 60-<70 years, 70-<80 years and ≥80 years showed upward trends (AAPC=11.657%, 14.031%, 10.734% and 5.300%, all P<0.05). A total of 3 739 deaths were reported, and the crude mortality, Chinese-standardized mortality and world-standardized mortality were 9.66/105, 5.23/105 and 3.71/105, respectively, showing upward trends (AAPC=8.458%, 3.620% and 3.602%, all P<0.05). The Chinese-standardized mortality of prostate cancer was higher in urban areas than in rural areas, and both showed upward trends (5.35/105 vs. 5.13/105; AAPC=3.183% and 3.962%, both P<0.05). The crude mortality of prostate cancer the groups ageds ≥80 years showed an upward trend (AAPC=7.482%, P<0.05).
Conclusions
From 2011 to 2023, the incidence and mortality of prostate cancer in Ningbo City showed upward trends. Special attention should be paid to urban residents, and prostate cancer screening should be strengthened among males aged 50 years and older.


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