1.Long-term survival outcomes and prognostic factors following radical resection of pancreatic body and tail cancer:a retrospective analysis of 992 patients
Dong XU ; Yang WU ; Kai ZHANG ; Nan LYU ; Qianqian WANG ; Pengfei WU ; Jie YIN ; Baobao CAI ; Guodong SHI ; Jianzhen LIN ; Yazhou WANG ; Lingdi YIN ; Zipeng LU ; Min TU ; Jianmin CHEN ; Feng GUO ; Jishu WEI ; Junli WU ; Wentao GAO ; Cuncai DAI ; Yi MIAO ; Kuirong JIANG
Chinese Journal of Surgery 2026;64(1):46-54
Objective:To investigate the survival outcomes and prognostic factors in patients undergoing radical resection for pancreatic body and tail cancer.Methods:A retrospective case series study was conducted on 992 patients who underwent radical resection for pancreatic body and tail cancer at the Pancreatic Center of the First Affiliated Hospital of Nanjing Medical University from January 2016 to June 2024. In this study, 577 (58.2%) were male and 415 (41.8%) were female,with an age of (65±9) years (range: 26 to 86 years). Follow-up continued until June 2024. Survival rates were estimated using the Kaplan-Meier method,and prognostic factors were identified using univariate and multivariate Cox proportional hazards models.Results:Among 992 patients,open surgery was the predominant approach (89.1%, 884/992), and radical antegrade modular pancreatosplenectomy (RAMPS) was performed in 317 patients (32.0%). Combined organ resection,venous resection,and arterial resection were performed in 23.5%, 9.3%,and 11.2% of patients,respectively. The rates of R0, R1-1 mm, and R1-direct resections were 49.8% (494/992),41.5% (412/992), and 8.7% (86/992),respectively. Stage ⅡB was the most common TNM stage (32.2%,319/992). A total of 801 patients (80.8%) received adjuvant chemotherapy. The median follow-up period was 32.0(8.8) months(range:3.2 to 105.3 months),during which 508 patients (51.2%) died. The overall median survival (OS) was 26.4 months,with 1-,3-, and 5-year survival rates of 79.0%,40.0%, and 29.0%, respectively. In the recent five years (from 2020 to 2024), the median OS improved significantly to 34.1 months compared to 20.0 months from 2016 to 2019 ( P<0.01). Histological subtype analysis showed that the median OS time was 26.7 months for pancreatic ductal adenocarcinoma (PDAC, n=855),58.9 months for invasive intraductal papillary mucinous carcinoma (IPMC, n=32),and 15.7 months for adenosquamous carcinoma of pancreas (ASCP, n=73) ( P=0.001). Among PDAC patients, adjuvant chemotherapy significantly improved survival (29.1 months vs. 14.4 months, P<0.01);in IPMC patients, adjuvant chemotherapy also extended survival (65.7 months vs. 58.9 months, P=0.047). Although ASCP patients receiving chemotherapy had a longer median OS time than those without (18.8 months vs. 8.9 months),the difference was not statistically significant ( P=0.151). Multivariate Cox regression analysis in PDAC patients indicated that adjuvant chemotherapy, R0 resection, T stage,N stage,and tumor differentiation were independent prognostic factors ( P<0.01). The median OS time by TNM stage was:not reached for stage ⅠA, 51.6 months for ⅠB, 25.5 months for ⅡA, 23.7 months for ⅡB, 23.0 months for Ⅲ, and 14.4 months for Ⅳ. The median OS time for R0,R1-1 mm,and R1-direct resections was 34.1,24.7,and 15.7 months,respectively ( P<0.01). Conclusion:Adjuvant chemotherapy,R0 resection,tumor stage,and differentiation are independent prognostic factors for pancreatic body and tail cancer.
2.Cuproptosis: A novel therapeutic target for non-small cell lung cancer
Dong DONG ; Yajie ZHANG ; Hecheng LI
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(05):826-832
Cuproptosis, recently defined as a unique form of cell death distinct from programmed cell death, is triggered by copper overload within mitochondria. Genes associated with cuproptosis have been found to correlate with tumorigenesis and tumor progression, making the targeting of cuproptosis pathways a promising direction for anti-tumor therapies. Copper ion carriers can transport copper ions into cells, inducing cuproptosis and laying the foundation for its application in cancer treatment. This article elaborates on the homeostasis of copper and the mechanisms related to cuproptosis, further clarifying the relationship between cuproptosis and lung cancer treatment targets. This review aims to summarize current progress in research related to cuproptosis and lung cancer, providing new theories and bases for the clinical treatment of lung cancer.
3.miR-433 inhibits cancer stem cell-like properties and enhances 5-FU sensitivity by targeting YAP1 in colorectal cancer HCT-116 cells
DONG Xinmin1 ; ZHENG Ti2△ ; LI Tiantian3 ; JUN Mei3 ; ZHANG Jian4
Chinese Journal of Cancer Biotherapy 2026;33(5):537-544
[摘 要] 目的:探讨miR-433对结直肠癌(CRC)细胞干细胞特性及5-氟尿嘧啶(5-FU)耐药性的影响,并阐明其潜在的分子调控机制。方法: 建立5-FU耐药的HCT-116细胞株。采用RT-qPCR和WB法检测miR-433及干细胞特性标志物(SOX2、OCT4、Nanog)的表达。通过双萤光素酶报告基因实验、免疫荧光和核质分离实验验证miR-433与靶基因YAP1的关系及YAP1对其亚细胞定位的影响。通过拯救实验确认miR-433/YAP1轴的功能。利用CCK-8法检测细胞对5-FU的敏感性。结果: 与亲代HCT-116/P细胞相比,耐药的HCT-116细胞株中miR-433表达显著下调(P < 0.05),而干细胞特性标志物(SOX2、OCT4、Nanog)表达上调(P < 0.05)。过表达miR-433能够抑制干细胞标志物的表达,并直接靶向YAP1,阻碍其核定位(P < 0.05)。恢复YAP1的表达能够部分逆转miR-433对干细胞标志物的抑制作用(P < 0.05)。功能上,上调miR-433显著增强了CRC细胞对5-FU的敏感性,而此效应同样可被YAP1的重新表达所削弱(P < 0.05)。结论: miR-433通过直接靶向YAP1通路,抑制CRC细胞的干细胞特性相关分子表型,并增强其对化疗药物的敏感性。miR-433/YAP1轴可能成为克服CRC 5-FU耐药的潜在新靶点。
4.Construction of the Diagnosis and Treatment System of "Sinew Prescription Correspondence" under the Guidance of Systematic Dialectical Sphygmology
Feng ZHANG ; Baoqiang DONG ; Xingxing LIN ; Yapeng LIU ; Lujia XIAO ; Bodong XING ; Yiyun CAO ; Wenhui ZHANG ; Wenqian QI
Journal of Traditional Chinese Medicine 2026;67(10):1038-1043
"Sinew prescription correspondence" is the principle of selecting prescriptions for channel sinew diseases. On the basis of the theory of syndrome differentiation and treatment, the pulse manifestation corresponds to the channel sinew syndrome, which can improve the flexibility and standardization of clinical prescriptions. From the perspective of systematic dialectical sphygmology, this paper explains the dialectical relationship between channel sinew theory and pulse body elements, pulse wall elements, pulse elements and blood flow elements, and clarifies the internal relationship between pulse manifestation and prescriptions at the level of channel sinew disease. The prescription is derived from the method, while the method is established with the syndrome, and the prescription is unified by the method. According to the theory of "sinew prescription correspondence", the treatment ideas of channel sinew diseases were analyzed from the perspective of channel sinew distribution, functional characteristics and structural changes. On this basis, the diagnosis of channel sinew disease and the application of prescriptions are expanded, and the research on the internal treatment and diagnosis mode of "pulse manifestation-channel sinew-zang fu (脏腑)" is prospected, so as to expand the differentiation and treatment methods of channel sinew theory.
5.Role of PI3K/Akt Pathway in Epirubicin Resistance in Triple-Negative Breast Cancer Explored Through Transcriptomic Analysis
Lingshan NAN ; Xiaomin WANG ; Xi ZUO ; Haiming LI ; Dong CHEN ; Xiaohui YIN ; Ganlin ZHANG
Cancer Research on Prevention and Treatment 2026;53(5):339-348
Objective To establish an epirubicin (EPI)-resistant murine triple-negative breast cancer (TNBC) (4T1/EPI) cell line and evaluate its biological characteristics and drug resistance. Methods The EPI-resistant cell line 4T1/EPI was developed through intermittent induction with gradually increasing EPI concentrations in vitro. Morphological changes were observed under an inverted microscope. Drug resistance index (MTT assay), cell doubling time (CCK-8 assay), and migration ability (wound healing assay) were evaluated. Western blot was used to detect the expression of drug resistance-related proteins. Transcriptome sequencing and KEGG pathway enrichment analysis were performed to identify the pathways and targets involved in EPI resistance, followed by experimental validation. Results The 4T1 cells eventually grew normally in a medium containing 100 ng/mL EPI, confirming the establishment of the 4T1/EPI resistant cell line. After stable resistance was acquired, morphological alterations were observed. Compared with their parental 4T1 cells, 4T1/EPI cells showed significantly prolonged doubling time (P<0.01) and enhanced migration ability (P<0.05). Expression levels of drug resistance-related proteins MDR1, MRP1 (P<0.01), and ABCG2 (P<0.05) were elevated in 4T1/EPI cells. In vivo models also demonstrated significant EPI resistance in 4T1/EPI tumors in terms of tumor weight and volume. Transcriptome sequencing highlighted the involvement of the PI3K/Akt signaling pathway and ABC transporter pathway. Validation experiments showed the upregulation of Erbb3, Egfr, PI3K, and Akt (P<0.05) and significant downregulation of Fgfr1 (P<0.01) in 4T1/EPI cells. Conclusion The EPI-resistant TNBC cell line 4T1/EPI was successfully established, exhibiting significant resistance in vitro and in vivo. The mechanism may involve the EPI-induced upregulation of Egfr and Erbb3, activating the PI3K/Akt pathway and subsequently enhancing ABC transporter expression.
6.Molecular Mechanism of Gypenoside L Inducing Ovarian Cancer Cell Apoptosis by Regulating NUF2 and Influencing Magnesium Homeostasis
Yang HONG ; Di ZHANG ; Yuanguang DONG ; Jiaxin WANG ; Lu PAN ; Lijiang ZHOU ; Mingdian YUAN ; Qun WANG ; Nan SONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):155-165
ObjectiveThis paper aims to investigate the role of NDC80 kinetochore complex component (NUF2) and magnesium homeostasis in ovarian cancer cell apoptosis, as well as the regulatory mechanism of gypenoside L (Gyp-L) on NUF2 and magnesium homeostasis. MethodsOvarian cancer OVCAR3 cells were divided into a blank control group, a low-concentration Gyp-L group (50 µmol·L-1), a high-concentration Gyp-L group (100 µmol·L-1), and a cisplatin (15 µmol·L-1) group. The migration, proliferation, and apoptosis capabilities of OVCAR3 cells were evaluated through cell scratch assays, clonal experiments, and terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining. Differentially expressed genes of ovarian cancer were screened by using the Gene Expression Omnibus (GEO) database. The interaction relationships of differentially expressed genes and proteins were analyzed via the Search Tool for Recurring Instances of Neighbouring Genes (STRING) database. The prognostic survival analysis was performed by using the Tumor Immune Estimation Resource (TIMER) database, and the differential expression levels of genes were validated with the Gene Expression Profiling Interactive Analysis (GEPIA) database. The mRNA expression levels of NUF2, magnesium homeostasis-related indicators, such as magnesium transporter 1 (MAGT1), non-imprinted in Prader-Willi/Angelman syndrome 1 (NIPA1), NIPA-like domain containing 1 (NIPAL1), as well as apoptosis-related indicators B cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in OVCAR3 cells, were detected by real-time quantitative polymerase chain reaction (Real-time PCR). The protein expression levels of NUF2, MAGT1, NIPA1, NIPAL1, Bcl-2, and Bax in OVCAR3 cells were quantitatively analyzed by ProteinSimple WES. A model of overexpression of NUF2 was constructed, and Gyp-L intervention was performed. The molecular mechanism by which Gyp-L induces ovarian cancer cell apoptosis by regulating NUF2 and influencing magnesium homeostasis was quantitatively analyzed and detected through cell cloning, TUNEL staining, Real-time PCR, and ProteinSimple WES. Finally, the Mg2+ content and protein synthesis efficiency were detected by immunofluorescence. ResultsGyp-L significantly inhibited the migration and proliferation capabilities of OVCAR3 cells and promoted their apoptosis (P<0.05). Overexpression of NUF2 markedly increased the expression levels of MAGT1, NIPA1, NIPAL1, and Bcl-2, while reducing the expression level of Bax (P<0.05). It also significantly elevated intracellular Mg2+ content and protein synthesis efficiency and simultaneously inhibited apoptosis (P<0.05). Gyp-L could reverse the magnesium homeostasis imbalance and apoptosis inhibition caused by the overexpression of NUF2, downregulating the expression levels of NUF2, MAGT1, NIPA1, NIPAL1, and Bcl-2 (P<0.05), while upregulating the expression level of Bax (P<0.05). ConclusionGyp-L can inhibit the occurrence of ovarian cancer, and its mechanism may involve inhibiting the expression of NUF2 to maintain magnesium homeostasis and inducing apoptosis of ovarian cancer cells.
7.Evaluation of CARIFS Score and Negative Antigen Conversion Rate of Qingxuan Daozhi Formula in Treatment of Influenza in Children (Heat Accumulation in Lung and Stomach Syndrome):A Multi-center Randomized Controlled Clinical Study
Jing WANG ; Liqun WU ; Tiegang LIU ; Yongning CAO ; Jing QIU ; Jing LI ; Huaqing TAN ; Ying ZHANG ; Xulei GOU ; Jia WANG ; Jing LI ; Haipeng CHEN ; Xueying QIN ; Yuanshuo TIAN ; Yang WANG ; Chen BAI ; Zhendong WANG ; Qianqian LI ; He YU ; Xueyan MA ; Fei DONG ; Lin JIANG ; Yingqi XU ; Jianping LIU ; Xiaohong GU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):188-196
ObjectiveThis paper aims to observe the syndrome improvement and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children (heat accumulation in the lung and stomach syndrome). MethodsThrough a multi-center randomized controlled methodology design,confirmed influenza cases were collected from October 2022 to April 2023 in the pediatrics department of eight hospitals,such as Dongfang Hospital of Beijing University of Chinese Medicine. A total of 180 children with influenza and heat accumulation in the lung and stomach syndrome conforming to the standard were recruited through the clinic. The sick children meeting the inclusion criteria were randomly divided into groups by a block-randomized method. The children in the experimental group were treated with Qingxuan Daozhi formula for five days,and those in the control group were treated with Oseltamivir Phosphate Granules for five days. The primary efficacy indicator was the negative conversion rate of influenza antigen detection. Secondary efficacy indicators were the Canadian acute respiratory illness and flu scale (CARIFS) and the incidence of complications,severe cases, and critical cases. Follow-up observation was conducted on the day of enrollment,48 hours after medication,72 hours after medication, and (6+1) d after medication. ResultsOne hundred and eighty participants were randomly assigned to the experimental group (90 cases) or the control group (90 cases). All participants were followed up during the study. Comparison of influenza antigen detection results in the primary efficacy indicators showed that the average time of negative influenza antigen conversion in the experimental group was (5.29±1.25) d,and that in the control group was (5.40±1.68) d,without a statistically significant difference. After five days of intervention,52 cases in the experimental group and 51 cases in the control group converted to negative,without a statistically significant difference. CARIFS score results in the secondary efficacy indicators showed that during 72 hours after intervention,there were statistically significant differences between the experimental group and the control group in three dimensions, including headache,muscle soreness, and the need for extra care (P<0.05). On the (6+1) days after the intervention,the differences in both the experimental group and the control group were statistically significant in 10 dimensions, including sore throat,bad sleep,uncomfortable feeling,poor spirit and fatigue,crying more than usual,the need for extra care,symptom,function,influence on parents,and total score (P<0.05). The comparison results within the group in the dimensional scores of symptom, function, and influence on parents,as well as the CARIFS total score showed that with the delay of follow-up time,scores of both groups decreased significantly,with a statistically significant difference (P<0.01). Inter-group comparison results showed that the mean score of the experimental group was higher than that of the control group at the time of enrollment. With the progress of intervention,the score of the experimental group was significantly decreased compared with that of the control group. At the end of follow-up,the mean score of the experimental group was lower than that of the control group,with no statistically significant difference. In terms of the incidence of complications,severe cases, and critical cases, there were no complications,severe cases, and critical cases in the two groups,without a statistically significant difference. ConclusionThe symptom improvement effect and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children (heat accumulation in the lung and stomach syndrome) are not inferior to Oseltamivir Phosphate granules, and children's acceptance is better. It can be more widely used in clinical treatment of influenza in children (heat accumulation in the lung and stomach syndrome).
8.Surveillance of Oncomelania hupensis snails following interruption of schistosomiasis transmission in Yunnan Province
Siqi NING ; Yi DONG ; Chunhong DU ; Lifang WANG ; Yun ZHANG ; Yuhe HE ; Hua JIANG ; Jiayu SUN ; Chunqiong CHEN ; Jiaqi YAN ; Jihua ZHOU ; Zongya ZHANG ; Hongqiong WANG ; Meifen SHEN ; Jing SONG
Chinese Journal of Schistosomiasis Control 2026;38(2):200-206
Objective To investigate the distribution characteristics of Oncomelania hupensis snails in Yunnan Province fol-lowing interruption of schistosomiasis transmission, so as to provide the evidence for assessing the risk of schistosomiasis transmission and scientifically formulating the schistosomiasis surveillance program. Methods According to the requirements of the National Schistosomiasis Surveillance Scheme (2020 Edition), O. hupensis snail surveillance data were collected from 18 schistosomiasis-endemic counties (cities, districts) in Yunnan Province from 2020 to 2024, including area of snail survey, area of snail habitats, area of re-emerging snail habitats, number of frames surveyed, number of frames with O. hupensis snails, number of O. hupensis snails captured, and number of living snails, and the occurrence of frames with snails and mean density of living snails were calculated. Changes in snail status over the 5-year period from 2020 to 2024 and the differences in snail distributions specified by epidemic intensity, environmental type, and vegetation type were analyzed. Results The areas of snail survey increased from 1 727.96 hm2 in 2020 to 3 894.45 hm2 in 2024 (peak) across 18 schistosomiasis-endemic counties (cities, districts) in Yunnan Province during the period from 2020 through 2024. The areas of snail habitats increased from 70.36 hm2 in 2020 to a peak in 2023 (172.04 hm2), followed by a reduction to 132.36 hm2 in 2024, and the areas of re-emerging snail habitats increased from 42.71 hm2 in 2020 to a peak in 2022 (78.43 hm2), followed by a reduction to 40.21 hm2 in 2024. The occurrence of frames with snails and mean density of living snails increased from 1.24% (3 025/244 404) and (0.033 2 ± 0.038 7) snails/0.1 m2 in 2020 to peaks at 2.03% (6 231/307 563) and (0.066 9 ± 0.068 4) snails/0.1 m2 in 2023, followed by reductions to 1.04% (5 829/559 941) and (0.032 6 ± 0.057 7) snails/0.1 m2 in 2024, respectively. There was a significant difference in the occurrence of frames with snails over the 5-year study period (χ2 = 1 962.95, P < 0.05), and the occurrence of frames with snails reduced by 48.71% in 2024 relative to in 2023 (χ2 = 1 411.05, P < 0.005); however, there was no significant difference in the mean density of living snails over the 5 years (H = 5.310, P > 0.05). There were significant differences in the occurrence of frames with snails (χ2 = 481.27, P < 0.05) and mean density of living snails (H = 6.872, P < 0.05) in schistosomiasis-endemic areas with different epidemic intensities. The occurrence of frames with snails (χ2 = 25.32 and 38.70, both P values < 0.017) and mean density of living snails (Z = 28.55 and 49.96, both P values < 0.017) were higher in schistosomiasis transmission-interrupted and eliminated areas with snails than in schistosomiasis-eliminated areas without snails, and the occurrence of frames with snails (χ2 = 453.54, P < 0.017) and mean density of living snails (Z = −56.97, P < 0.017) were higher in schistosomiasis-eliminated areas with snails than in schistosomiasis transmission-interrupted areas with snails. O. hupensis snails were mainly distributed in paddy fields, dry farmlands and ditches; however, the occurrence of frames with snails (13.40%, 424/3 164) and mean density of living snails [(0.252 8 ± 0.158 7) snails/0.1 m2] were higher in ponds/weirs than in other types of environments (both P values < 0.05). Rice, dry farmland crops and weeds were main vegetations in which O. hupensis snails were distributed, and the occurrence of frames with snails (2.29%, 7 111/310 140) and mean density of living snails [(0.072 3 ± 0.018 9) snails/0.1 m2] were higher in weeds than in other types of environments (both P values < 0.05). Conclusions O. hupensis snails have been effectively controlled in Yunnan Province following implementation of integrated schistosomiasis control measures; however, there are still risk factors for schistosomiasis transmission, including reduced attention to schistosomiasis control and snail re-emergence. Improved control efforts and surveillance system construction and timely identification of risk factors of snail status and timely management are recommended to ensure the achievement of the target of schistosomiasis elimination as scheduled.
9.Research Progress on Animal Models of Systemic Sclerosis with Microvascular Involvement
Haochen HUANG ; Lingling ZHANG ; Dong XU
Medical Journal of Peking Union Medical College Hospital 2026;17(3):834-841
The early angiopathy of systemic sclerosis (SSc) is a key hub connecting inflammation and fibrosis. The animal models that simulate the pathological characteristics of SSc are of great significance to study the etiology, pathogenesis and treatment of SSc with vascular involvement. This review of the characteristics of different animal models of SSc with vascular involvement provides a reference for relevant researchers to select appropriate models in the process of exploring the pathogenesis of SSc with vascular involvement.
10.Qiangjing Tablets Regulate CDK4-E2F Signaling Pathway to Delay Aging of Leydig Cells and Testicular Tissue in Rats
Xiucheng LAN ; Meijing WANG ; Jingyi ZHANG ; Junjun LI ; Liang DONG ; Xujun YU ; Fang YANG ; Degui CHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):328-336
ObjectiveTo reveal the molecular mechanism by which the traditional Chinese medicine compound prescription Qiangjing tablets regulate the aging of the testicular tissue and Leydig cells in rats through the cyclin-dependent kinase 4 (CDK4)-early 2 factor (E2F) signaling pathway. MethodsFor the cell experiment, 2-month-old SPF-grade SD male rats were selected and randomly assigned into a blank control group (administrated with an equal volume of 0.9% sodium chloride injection) and a Qiangjing tablets group (20 rats in each group) according to body weight. The Leydig cell model of aging was established by treatment of TM3 cells with 100 μmol·L-1 H2O2, and the modeling performance was evaluated based on the levels of p16 and p21 determined by Western blot. The antioxidant NAC (1 mmol·L-1) was used as the positive control for eliminating reactive oxygen species (ROS). Cells were intervened with Qiangjing tablets-containing serum at low (2.5%), medium (5%), and high (10%) concentrations. The testosterone level in the cell supernatant was determined by enzyme-linked immunosorbent assay (ELISA), and the protein levels of CDK4, E2F1, and E2F2 were analyzed by Western blot. In the animal experiment, 19-month-old naturally aging rats were used as the model group, and 2-month-old rats as the young control group. The positive control group was subcutaneously injected with 5.21 mg·kg-1·d-1 testosterone propionate. Qiangjing tablets were administered by gavage at low, medium, and high doses of 0.72, 1.44, 2.88 g·kg-1·d-1, respectively. The general conditions of rats were observed, and the protein levels of CDK4, E2F1, and E2F2 in the testicular tissue were determined by Western blot. ResultsIn the cell experiment, compared with the blank control group, the model group showed upregulated expression of CDK4 and E2F1 (P<0.05) and slightly downregulated expression of E2F2. Compared with that in the model group, the expression of CDK4 was upregulated in the NAC group and the low-dose Qiangjing tablets group (P<0.05), slightly upregulated in the medium-dose Qiangjing tablets group, and downregulated in the high-dose Qiangjing tablets group (P<0.05). The NAC group showed downregulated expression of E2F1 (P<0.05) and E2F2, and the low-, medium-, and high-dose Qiangjing tablets groups showed downregulated expression of both E2F1 and E2F2 (P<0.05). Compared with that in the NAC group, the expression of CDK4 was upregulated in the low-dose Qiangjing tablets group and downregulated in the medium-dose and high dose (P<0.05) groups. The expression of E2F1 was down-regulated in all the three dose groups, with statistically significance in the high dose group (P<0.05), and that of E2F2 were downregulated in all the three dose groups (P<0.05). In the animal experiment, compared with the young control group, the model group exhibited downregulated expression of CDK4 (P<0.05) and slightly upregulated expression of E2F1 and E2F2. Compared with that in the model group, the expression of CDK4 decreased in the testosterone propionate group and the low-dose Qiangjing tablets group (P<0.05) but increased in the medium-dose (P<0.05) and high-dose groups. In addition, the expression of E2F1 decreased (P<0.05), and that of E2F2 was slightly elevated. Compared with that in the NAC group, CDK4 expression was elevated in the Qiangjing tablets groups, with statistical significance in the medium- and high-dose groups (P<0.05). Similarly, the E2F1 expression was also upregulated in the Qiangjing tablets groups, with statistical significance in the medium-dose group (P<0.05). The expression of E2F2 was downregulated in all the Qiangjing tablets groups. ConclusionQiangjing tablets delay the aging process of Leydig cells and testicular tissue by up-regulating the expression of CDK4 and lowering the levels of E2F1 and E2F2.

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