1.Construction and preparation of virus-like particle expression vector for yellow fever virus
Miaomiao LYV ; Chunying GAO ; Yunfeng LI ; Zhiying JIN ; Min WAN ; Jie TIAN ; Weina KANG
Journal of Public Health and Preventive Medicine 2026;37(5):12-16
Objective To prepare virus-like particles (VLPs) for yellow fever virus (YFV) using the pACYCDuet-1 single-plasmid dual expression system. Methods Gene fragments including NS1, NS5, and 5' untranslated region (UTR) and 3' UTR fragment of YFV were cloned into the expression plasmid pACYCDuet-1-MS2-TMpac (carrying MS2 coat protein gene) and transformed into E. coli BL21(DE3) for expression. VLPs were observed by TEM. After DNase I/RNase A digestion, RNA was extracted and transcribed into cDNA, which was subjected to fluorescence quantitative PCR detection. Results According to SN/T 3560-2013, double digestion and colony PCR of pACYCDuet-1-MS2-TMpac-YFV both produced expected bands. SDS-PAGE showed a target band at 25-35 kDa. RT-PCR amplified VLPs after enzyme digestion, and TEM revealed VLPs with clear edges and translucent interiors. Thus, the plasmid was successfully constructed, and the phage coat protein packaged YFV genes with nuclease resistance. Conclusion VLPs containing specific gene fragments of yellow fever virus have been successfully prepared. These nuclease-resistant VLPs can serve as safe and stable quality control materials to ensure the reliability of laboratory detection for YFV.
2.Buzhong Yiqitang Regulates Endoplasmic Reticulum Stress via Nrf2/ROS/PERK/CHOP Signaling Pathway to Attenuate Cisplatin Resistance in NSCLC
He LI ; Yuetong LIU ; Jingyi HUANG ; Qirui MU ; Chunying LIU ; Yuan GAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):79-89
ObjectiveTo explore the molecular mechanism of Buzhong Yiqitang in attenuating cisplatin resistance of non-small cell lung cancer (NSCLC) cells (A549/DDP) by regulating endoplasmic reticulum stress (ERS) via the nuclear factor E2-related factor 2 (Nrf2)/reactive oxygen species (ROS)/double-stranded RNA-activated protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK)/CCAAT enhancer-binding protein homologous protein (CHOP) signaling pathway. MethodsSprague Dawley
3.Buzhong Yiqitang Induces Ferroptosis by Regulating PCBP1 to Attenuate Cisplatin Resistance in Non-small Cell Lung Cancer
Yuetong LIU ; He LI ; Qirui MU ; Jingyi HUANG ; Haoran CAI ; Chunying LIU ; Yuan GAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):90-97
ObjectiveTo explore the molecular mechanism of Buzhong Yiqitang in attenuating cisplatin resistance in non-small cell lung cancer (NSCLC) by inducing ferroptosis via poly(rC)-binding protein 1 (PCBP1). MethodsThe serum containing Buzhong Yiqitang was prepared and cisplatin-resistant human non-small cell lung cancer (NSCLC) cells (A549/DDP) were cultured and randomly grouped as follows: Blank (10% blank serum), model (10% blank serum+20 mg·L-1 cisplatin), Buzhong Yiqitang (10% serum containing Buzhong Yiqitang+20 mg·L-1 cisplatin), Fe-1 (10% blank serum+20 mg·L-1 cisplatin+5 μmol·L-1 Fe-1), and Buzhong Yiqitang+Fe-1 (10% serum containing Buzhong Yiqitang+20 mg·L-1 cisplatin+5 μmol·L-1 Fe-1). Firstly, PCR Array was used to screen ferroptosis-related genes regulated by Buzhong Yiqitang, and PCBP1 was identified as the target for studying the attenuation of cisplatin resistance by Buzhong Yiqitang. Subsequently, the median inhibitory concentration (IC50) of cisplatin in each group was determined by the cell counting kit-8 (CCK-8) method and the resistance index (RI) was calculated. The ultrastructure of A549/DDP cells in each group was observed by transmission electron microscopy. The protein levels of PCBP1 and glutathione peroxidase 4 (GPX4) were determined by Western blot. The lipid reactive oxygen species (ROS) content in each group was determined by the C11-BODIRY 581/591 fluorescence probe. The ferrous ion assay kit was used to measure the ferrous ion content in each group. The malondialdehyde (MDA) assay kit was used to determine the MDA content in each group. ResultsCompared with model group, the IC50 of cisplatin and the RI of A549/DDP cells decreased in the Buzhong Yiqitang group (P<0.05) but increased in the Fe-1 group (P<0.05). The IC50 of cisplatin and the RI of A549/DDP cells in the Buzhong Yiqitang+Fe-1 group were lower than those in the Fe-1 group (P<0.05). Compared with the model group, the Buzhong Yiqitang group showed obvious mitochondrial ferroptosis, while the mitochondrial damage became less obvious after Fe-1 treatment. Compared with that in the Fe-1 group, the mitochondrial ferroptosis was aggravated after the intervention with Buzhong Yiqitang. Compared with blank group, the model group showed down-regulated expression levels of PCBP1 and GPX4 (P<0.05) and increased content of lipid ROS, ferrous ions, and MDA (P<0.05) in A549/DDP cells. Compared with model group, the Buzhong Yiqitang group showed down-regulated expression levels of PCBP1 and GPX4 (P<0.05) and increased content of lipid ROS, ferrous ions, and MDA (P<0.05), while the Fe-1 group showed up-regulated expression levels of PCBP1 and GPX4 (P<0.05) and reduced content of lipid ROS, ferrous ions, and MDA (P<0.05). Compared with the Fe-1 group, the Buzhong Yiqitang+Fe-1 group showed down-regulated expression levels of PCBP1 and GPX4 and increased content of lipid ROS, ferrous ions, and MDA (P<0.05). ConclusionBuzhong Yiqitang attenuated cisplatin resistance in NSCLC by regulating PCBP1 to induce ferroptosis.
4.Traditional Chinese Medicine Ameliorates Tumor Chemotherapy Resistance: A Review
Jingyi HUANG ; Yuetong LIU ; He LI ; Qirui MU ; Chenyi LI ; Chunying LIU ; Yuan GAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):105-116
In the process of tumor chemotherapy, the emergence of multi-drug resistance (MDR) has always been a thorny problem, which is a result of the joint action of the host, tumor cells, and the immune microenvironment. Tumor cells can escape the toxicity of chemotherapeutic drugs through multiple pathways, being easy to produce drug resistance. MDR greatly restricts the effect of chemotherapeutic drugs on tumor cells and affects their therapeutic effects. Traditional Chinese medicine (TCM) has the unique advantages of multi-target, multi-pathway and individualized treatment. The TCM treatment of tumors emphasizes regulating Yin and Yang, as well as reinforcing healthy Qi and dispelling pathogen. In recent years, TCM has demonstrated remarkable efficacy in the treatment of tumors and the amelioration of multi-drug resistance. TCM not only can target the phenomenon of MDR but also greatly weakens the side effects of the patients after the chemotherapy, thus improving the survival quality and rate of the patients. Accordingly, many patients adopt TCM as an adjuvant therapy during or after chemotherapy. The binding of TCM to targets can reverse the drug resistance of various tumors, which has become an emerging research highlight. From the regulatory mechanism of TCM on MDR of tumors, this paper introduces the mechanisms by which tumor cells continue to grow, proliferate, and metastasize by adjusting the intracellular drug concentration, altering or utilizing the tumor microenvironment, and affecting the cell death mode to achieve the resistance to chemotherapeutic drugs. In this regard, the active ingredients and compound prescriptions of TCM can increase the sensitivity of chemotherapeutic drugs by down-regulating drug transporters, improving the tumor microenvironment, and modulating the drug resistance pathways associated with apoptosis, autophagy, ferroptosis, or pyroptosis. The aim of this paper is to explore more clinical practical value of TCM in the treatment of tumors and provide exploratory ideas and a theoretical basis for the future research on tumors and MDR.
5.Toxicological evaluation of aristolochic acid II following single and repeated oral administration over a 24-week period
Yan YI ; Chunying LI ; Yong ZHAO ; Jingzhuo TIAN ; Yuan WANG ; Yushi ZHANG ; Suyan LIU ; Chen PAN ; Lianmei WANG ; Shuangrong GAO ; Jianyin HAN ; Zhong XIAN ; Chenyue LIU ; Dunfang WANG ; Jing MENG ; Meiting LIU ; Aihua LIANG
Science of Traditional Chinese Medicine 2025;3(4):366-377
Background: Aristolochic acid II (AAII), a major nephrotoxic and carcinogenic component of aristolochic acids (AAs), has been less studied compared with its well-characterized analog, aristolochic acid I (AAI). Although AAs are known to induce carcinogenesis via DNA adduct formation, the toxicity mechanisms, environmental prevalence, and long-term health impacts of AAII remain poorly understood. Objective: This study aimed to systematically evaluate AAII’s acute and chronic toxicity, carcinogenic mechanisms, and environmental exposure patterns using integrated murine models and phytochemical analyses to clarify its toxicological profile and associated health risks. Methods: C57BL/6J mice were used in the following experiments: (1) determination of AAII content in 3 commonly used Aristolochia medicinal materials via liquid chromatography-mass spectrometry/mass spectrometry; (2) acute toxicity testing with single doses of 10, 20, or 40 mg/kg; and (3) chronic exposure with 1 or 10 mg/kg administered every other day for 24 weeks, followed by 21 to 40 weeks of postexposure monitoring. Histopathological examination, whole-exome sequencing, biochemical assays, and micronucleus tests were performed to assess multi-organ damage, tumorigenesis, genomic mutation signatures, and direct clastogenicity. Phytochemical analyses were used to evaluate environmental distribution. Results: (1) A single 40 mg/kg dose of AAII induced dose-dependent renal tubular degeneration without hepatotoxicity; (2) the 10 mg/kg group showed significant mortality (20%), tumor incidence (33.3%, primarily forestomach and bladder transitional cell carcinomas), persistent renal interstitial fibrosis, and subclinical hepatic injury. Chronic exposure to 1 mg/kg still induced 13.3% mortality and 15.5% tumor incidence over a 64-week period; (3) whole-exome sequencing revealed a predominance of C>T mutations and pathway enrichment in chemical carcinogenesis and cytochrome P450-mediated metabolism, indicating reactive metabolite-driven mechanisms distinct from classical AA-DNA adducts; and (4) no histopathological changes were observed in nontarget organs (brain, heart, and testes), and micronucleus assays confirmed the absence of direct clastogenicity. Conclusion: This study highlights the delayed carcinogenic risks of low-dose chronic AAII exposure and emphasizes the need to update regulatory frameworks to ensure the safe use of aristolochiaceae-containing herbal products.
6.Mid-and long-term effect of Kegel training combined with Pilates training on urinary control recovery in pa-tients with post-prostatectomy incontinence with different body mass index
Di AN ; Jianxia WANG ; Fan ZHANG ; Huafang JING ; Yi GAO ; Huiling CONG ; Guodong SU ; Miao YE ; Chunying HU ; Juan WU ; Limin LIAO
Chinese Journal of Rehabilitation Theory and Practice 2025;31(8):972-978
Objective To observe the mid-and long-term effects of Kegel training combined with Pilates training on urinary conti-nence recovery in different body mass index(BMI)male patients with urinary incontinence after prostatectomy.Methods From May,2023 to June,2024,48 patients in Beijing Bo'ai Hospital were recruited and divided into group A(<25 kg/m2,n=15),group B(25 to 30 kg/m2,n=18)and group C(>30 kg/m2,n=15)according to their BMI.All the groups performed Kegel training combined with Pilates training for two months,and followed up at six months from baseline.They were evaluated with one hour pad test,the number of daily urinary incontinence,In-ternational Consultation on Incontinence Questionnaire-Short Form(ICIQ-SF)and modified Oxford Rating Scale before treatment,and four weeks,eight weeks and six months after treatment.Results The intra-group effect,the inter-group effect and interaction effect were significant in the results of one hour pad test and the daily number of urinary incontinence(F>2.955,P<0.05).Post Hoc test showed that they were worse in group C than in groups A and B(P<0.05),and the number of daily urinary incontinence was more in group B than in group A(P<0.05).There was significant difference in the scores of ICIQ-SF and modified Ox-ford Rating Scale among groups in different time points after treatment(Z>10.476,P<0.05)except the score of ICIQ-SF four weeks after treatment(P>0.05),and they were the worst in group C.BMI(group A=1,group B=2,group C=3)was correlated with the results of one hour pad test(r=0.79,P<0.001),the number of daily uri-nary incontinence(r=0.68,P<0.001),and the scores of ICIQ-SF(r=0.68,P<0.001)and modified Oxford Rating Scale(r=-0.47,P=0.001)six months after treatment.Conclusion Kegel training combined with Pilates training could improve the urinary control in patients with urinary in-continence after prostatectomy.The decrease of BMI can promote the recovery of urinary control,and improve the symptoms of later urinary incontinence in mid-and long-term.
7.Mid-and long-term effect of Kegel training combined with Pilates training on urinary control recovery in pa-tients with post-prostatectomy incontinence with different body mass index
Di AN ; Jianxia WANG ; Fan ZHANG ; Huafang JING ; Yi GAO ; Huiling CONG ; Guodong SU ; Miao YE ; Chunying HU ; Juan WU ; Limin LIAO
Chinese Journal of Rehabilitation Theory and Practice 2025;31(8):972-978
Objective To observe the mid-and long-term effects of Kegel training combined with Pilates training on urinary conti-nence recovery in different body mass index(BMI)male patients with urinary incontinence after prostatectomy.Methods From May,2023 to June,2024,48 patients in Beijing Bo'ai Hospital were recruited and divided into group A(<25 kg/m2,n=15),group B(25 to 30 kg/m2,n=18)and group C(>30 kg/m2,n=15)according to their BMI.All the groups performed Kegel training combined with Pilates training for two months,and followed up at six months from baseline.They were evaluated with one hour pad test,the number of daily urinary incontinence,In-ternational Consultation on Incontinence Questionnaire-Short Form(ICIQ-SF)and modified Oxford Rating Scale before treatment,and four weeks,eight weeks and six months after treatment.Results The intra-group effect,the inter-group effect and interaction effect were significant in the results of one hour pad test and the daily number of urinary incontinence(F>2.955,P<0.05).Post Hoc test showed that they were worse in group C than in groups A and B(P<0.05),and the number of daily urinary incontinence was more in group B than in group A(P<0.05).There was significant difference in the scores of ICIQ-SF and modified Ox-ford Rating Scale among groups in different time points after treatment(Z>10.476,P<0.05)except the score of ICIQ-SF four weeks after treatment(P>0.05),and they were the worst in group C.BMI(group A=1,group B=2,group C=3)was correlated with the results of one hour pad test(r=0.79,P<0.001),the number of daily uri-nary incontinence(r=0.68,P<0.001),and the scores of ICIQ-SF(r=0.68,P<0.001)and modified Oxford Rating Scale(r=-0.47,P=0.001)six months after treatment.Conclusion Kegel training combined with Pilates training could improve the urinary control in patients with urinary in-continence after prostatectomy.The decrease of BMI can promote the recovery of urinary control,and improve the symptoms of later urinary incontinence in mid-and long-term.
8.Buzhong Yiqitang Regulates Apoptosis via Nrf2/ROS Pathway to Alleviate Cisplatin Resistance in A549/DDP Cells
Qirui MU ; He LI ; Yuetong LIU ; Yuan GAO ; Chunying LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(1):95-102
ObjectiveTo explore the mechanism of Buzhong Yiqitang-containing serum in alleviating the cisplatin resistance in human non-small cell lung cancer (A549/DDP) cells via regulating the nuclear factor E2-related factor 2 (Nrf2)/reactive oxygen species (ROS) signaling pathway. MethodThe serum containing Buzhong Yiqitang was prepared and A549/DDP cells were cultured and randomly grouped: blank (10% blank serum), cisplatin (10% blank serum+20 mg·L-1 cisplatin), Buzhong Yiqitang (10% Buzhong Yiqitang-containing serum+20 mg·L-1 cisplatin), ML385 (10% blank serum+5 μmol·L-1 ML385+20 mg·L-1 cisplatin), Buzhong Yiqitang+ML385 (10% Buzhong Yiqitang-containing serum+5 μmol·L-1 ML385+20 mg·L-1 cisplatin), tertiary butylhydroquinone (TBHQ) (10% blank serum+5 μmol·L-1 TBHQ+20 mg·L-1 cisplatin), and Buzhong Yiqitang+TBHQ (10% Buzhong Yiqitang-containing serum+5 μmol·L-1 TBHQ+20 mg·L-1 cisplatin). The median inhibitory concentration (IC50) of cisplatin in each group was determined by the cell counting kit-8 (CCK-8) method and the resistance index (RI) was calculated. The apoptosis rate was detected by flow cytometry. The ROS content of each group was determined with the DCFH-DA fluorescence probe. Western blot was employed to determine the protein levels of Nrf2, cleaved cysteinyl aspartate-specific protease-3 (cleaved Caspase-3), cytochrome C (Cyt C), and B-cell lymphoma-2 (Bcl-2). ResultCompared with those in the cisplatin group, the IC50 and RI of A549/DDP cells to cisplatin in Buzhong Yiqitang, ML385, and Buzhong Yiqitang+ML385 groups decreased (P˂0.05). Compared with the blank group, the cisplatin, Buzhong Yiqitang, ML385, and Buzhong Yiqitang+ML385 groups showed increased apoptosis rate of A549/DDP cells (P˂0.05). Compared with the blank group, cisplatin promoted the expression of Nrf2 (P˂0.05). Compared with the cisplatin group, Buzhong Yiqitang, ML385, and Buzhong Yiqitang+ML385 inhibited the expression of Nrf2 (P<0.05), elevated the ROS level (P˂0.05), up-regulated the protein levels of cleaved Caspase-3 and Cyt C, and down-regulated the protein level of Bcl-2 (P<0.05), which were the most significant in the Buzhong Yiqitang+ML385 group. Compared with the cisplatin group, the TBHQ group showed increased IC50 and RI of cisplatin (P<0.05), decreased apoptosis rate of A549/DDP cells (P<0.05), up-regulated protein levels of Nrf2 and Bcl-2 (P<0.05), lowered level of ROS (P˂0.05), and down-regulated protein levels of cleaved Caspase-3 and Cyt C (P<0.05). Compared with the TBHQ group, Buzhong Yiqitang+TBHQ decreased the IC50 and RI of cisplatin in A549/DDP cells (P<0.05), increased the apoptosis rate (P<0.05), down-regulated the protein levels of Nrf2 and Bcl-2 (P<0.05), increased ROS (P˂0.05), and up-regulated the protein levels of cleaved Caspase-3 and Cyt C (P<0.05). ConclusionBuzhong Yiqitang induced apoptosis by inhibiting Nrf2/ROS pathway to alleviate cisplatin resistance in A549/DDP cells.
9.Shear wave viscoelastography for differentiating lung peripheral inflammatory masses and malignant tumors
Jiling WEI ; Chunying LI ; Han YUAN ; Hengfei CHEN ; Yong GAO ; Xinhong LIAO
Chinese Journal of Medical Imaging Technology 2024;40(10):1524-1528
Objective To observe the value of shear wave viscoelastography(SWV)for differentiating lung peripheral inflammatory masses and malignant tumors.Methods Conventional gray-scale ultrasound and SWV were prospectively performed in 70 patients with lung peripheral inflammatory mass or malignant tumor.The patients were divided into malignant group(n=42)and inflammatory group(n=28)according to pathological results.Clinical and ultrasonic data,including the maximum diameter of lesions,the mean Young's modulus(Emean),mean viscosity(Vmean),and mean dispersion slope(Dmean)were compared between groups.Receiver operating characteristic curves of ultrasonic parameters being significantly different between groups were drawn,and area under the curves(AUCs)were calculated to evaluate the efficacy of each parameter for differentiating lung peripheral inflammatory mass or malignant tumor.Results In malignant group,the maximum diameter and Emean of lesions were both higher,while Vmean and Dmean of lesions were both lower than those in inflammatory group(all P<0.05).Vmean and Dmean of lesions had moderately/good efficacy for differentiating lung peripheral inflammatory mass or malignant tumor(AUC=0.843,0.866),both better than that of conventional ultrasound and Emean(AUC=0.673,0.685)(all P<0.05).The combination of Emean,Vmean and Dmean had good efficacy for differentiating lung peripheral inflammatory masses and malignant tumors,with AUC of 0.874.Conclusion The viscous parameters of SWV could effectively differentiating lung peripheral inflammatory masses and malignant tumors.
10.Comparison of the efficacy of fractional CO 2 laser combined with topical delivery of fluorouracil versus compound betamethasone injections in the treatment of vitiligo: a clinical observation
Qian ZHANG ; Jin′e ZHANG ; Sen GUO ; Pu SONG ; Lin GAO ; Chunying LI
Chinese Journal of Dermatology 2024;57(1):34-38
Objective:To compare the efficacy of fractional CO 2 laser combined with topical delivery of fluorouracil versus compound betamethasone injections in the treatment of vitiligo. Methods:Clinical data were collected from 94 patients with localized, non-segmental, and stable vitiligo, who received fractional CO 2 laser combined with drug delivery at the Cosmetological Center, Xijing Hospital, Air Force Medical University from October 2018 to May 2023, and were retrospectively analyzed. Among them, there were 40 cases in the fractional CO 2 laser combined with fluorouracil injection group, and 54 cases in the fractional CO 2 laser combined with compound betamethasone injection group. All the patients received the above treatment once a month for 5 sessions. A 4-level grading scale was used to evaluate the pigmentation improvement, and the clinical efficacy and safety of the two therapeutic regimens were compared. Comparisons between groups were performed using chi-square test, Fisher′s exact test, and t test. Results:In the fractional CO 2 laser combined with fluorouracil injection group, there were 22 males and 18 females, their ages were 21.95 ± 12.88 years, and the disease duration was 25.46 ± 11.42 months; in the fractional CO 2 laser combined with compound betamethasone injection group, there were 36 males and 18 females, their ages were 22.26 ± 8.79 years, and the disease duration was 26.51 ± 12.81 months. One month after the first treatment, no significant difference was observed in the efficacy between the two groups ( χ2 = 1.39, P = 0.238). One month after the fifth treatment, 2 (5.0%) patients showed an excellent response, 4 (10.0%) showed a good response, 12 (30.0%) showed a mild response, and 22 (55.0%) showed a poor response in the fractional CO 2 laser combined with fluorouracil injection group; in the fractional CO 2 laser combined with compound betamethasone injection group, 8 (14.8%) patients showed a good response, 8 (14.8%) showed a mild response, and 38 (70.4%) showed a poor response; there was no significant difference in the efficacy between the two groups after 5 sessions of treatment ( χ2 = 2.35, P = 0.125). After either 1 or 5 sessions of treatment, there were no significant differences in the efficacy for lesions on the face and neck, trunk and limbs, hands and feet between the two therapeutic regimens (all P > 0.05). Comparisons of the efficacy for skin lesions on different body sites showed that one session of the fractional CO 2 laser combined with fluorouracil injection was more effective for the treatment of skin lesions on the face and neck compared with those on the hands and feet ( P = 0.039) ; after 5 sessions of treatment, the two therapeutic regimens both showed better efficacy for facial skin lesions compared with hand and foot skin lesions ( P = 0.005, 0.049). There was no significant difference in the occurrence of adverse reactions such as pigmentation, infection and scarring between the two groups. Conclusion:The fractional CO 2 laser combined with topical delivery of fluorouracil and compound betamethasone injections showed similar efficacy and safety in the treatment of vitiligo, and both can be used as treatment options for vitiligo.


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