1.Construction and characterization of recombinant human coagulation factor Ⅶ stable transfected cell lines
Xiaoxiao LI ; Jiabin CHEN ; Jiajun LIU ; Zhifei ZHANG ; Sen ZOU ; Lihua ZHU ; Zhaoyong YANG
Acta Universitatis Medicinalis Anhui 2026;61(1):16-22
ObjectiveTo construct a stable monoclonal human embryonic kidney 293 (HEK293) cell line expressing recombinant human coagulation factor Ⅶ (rhFⅦ) and evaluate the expression level and procoagulant bioactivity of rhFⅦ. MethodsThe plasmid pCDNA3.1-EGFP-FⅦ was transfected into HEK293 cells to verify the effectiveness of the transfection system. The plasmid pCDNA3.1-FⅦ was transfected into HEK293 cells, and monoclonal stable transfected cell lines were selected using geneticin (G418). The transcription of the FⅦ gene was identified by reverse transcription polymerase chain reaction (RT-PCR). The expression level of rhFⅦ in the supernatant of the monoclonal stable transfected cell line was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. The concentration of rhFⅦ was determined by enzyme-linked immunosorbent assay (ELISA), and the procoagulant activity of rhFⅦ was measured by human coagulation factor Ⅶ potency assay. ResultsHEK293 cells transfected with pcDNA3.1-EGFP-FⅦ showed green fluorescence, indicating that rhFⅦ was successfully expressed in the supernatant of HEK293 cells after transient transfection with pcDNA3.1-FⅦ. The monoclonal stable transfected cell line was obtained by G418 screening. RT-PCR identified that the FⅦ gene was integrated into the genome of the monoclonal stable transfected cell line. The cell viability was good as detected by Cell Counting Kit-8, and a single band of rhFⅦ was obtained by purification of the cell supernatant. The highest rhFⅦ expression was (1.27±0.09) mg/L, and the highest procoagulant activity was (380.29±13.80)%. ConclusionThe monoclonal HEK293 cell lines which can express rhFⅦ protein efficiently and stably with excellent procoagulant bioactivity is successfully screened.
2.The effects of galangin on the apoptosis and autophagy of gastric cancer NCI-N87 cells through regulating the AMPK/mTOR/ULK1 signaling pathway
GUO Fang ; CHEN Wei ; LIU Meng ; ZOU Yanli ; TIAN Xia
Chinese Journal of Cancer Biotherapy 2026;33(1):59-65
[摘 要] 目的:探讨高良姜素(Gal)调控AMPK/mTOR/ULK1信号通路对胃癌细胞凋亡和自噬的影响及其机制。方法:将胃癌NCI-N87细胞分为对照组、多索吗啡(DM)组、Gal低剂量(Gal-L)组、Gal高剂量(Gal-H)组、Gal-H + DM组。采用MTT法、流式细胞术、划痕愈合实验和Transwell实验分别检测各组细胞的增殖、凋亡、迁移和侵袭能力,WB法检测PCNA、C-caspase-3、免疫逃逸相关蛋白(B7H1)、EMT和AMPK/mTOR/ULK1信号通路蛋白的表达水平。建立裸鼠NCI-N87细胞移植瘤模型,观察Gal和5-FU对移植瘤的抑制效果。结果:与对照组比较,DM组NCI-N87细胞增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著升高(均P < 0.05),细胞凋亡率、C-caspase-3、E-cadherin、LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著降低(均P < 0.05);Gal-L组和Gal-H组NCI-N87细胞的增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著降低(均P < 0.05),细胞凋亡率、C-caspase-3、E-cadherin、LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著升高(均P < 0.05);DM可部分逆转Gal对NCI-N87细胞恶性生物学行为的抑制作用(P < 0.05);与对照组比较,Gal组和5-FU组裸鼠移植瘤体积和质量均显著降低,肿瘤组织细胞凋亡率显著升高(P < 0.05)。结论:Gal可促进胃癌NCI-N87细胞自噬和凋亡,抑制其增殖、迁移和侵袭,可能与激活AMPK/mTOR/ULK1信号通路有关。
3.Yimei Baijiang Formula Treats Colitis-associated Colorectal Cancer in Mice via NF-κB Signaling Pathway
Qian WU ; Xin ZOU ; Chaoli JIANG ; Long ZHAO ; Hui CHEN ; Li LI ; Zhi LI ; Jianqin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):119-130
ObjectiveTo explore the effects of Yimei Baijiang formula (YMBJF) on colitis-associated colorectal cancer (CAC) and the nuclear factor kappaB (NF-κB) signaling pathway in mice. MethodsSixty male Balb/c mice of 4-6 weeks old were randomized into 6 groups: Normal, model, capecitabine (0.83 g
4.Animal Model of Chronic Obstructive Pulmonary Disease and Intervention Effect of Traditional Chinese Medicine: A Review
Jiyu ZOU ; Lijian PANG ; Tianjiao WANG ; Ningzi ZANG ; Zhongxue ZHAO ; Yongming LIU ; Qi SI ; Tianya CAO ; Xuenan MA ; Ying WANG ; Jiaran WANG ; Xiaodong LYU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):294-303
Chronic obstructive pulmonary disease (COPD), as one of the three major causes of death, is a complex systemic disease with high prevalence, high mortality, high disability, frequent acute exacerbations, and a variety of pulmonary complications. The pathogenesis is complex. Western medicine has no effective specificity scheme for a complete cure. However, multiple-component and multiple-target characteristics of traditional Chinese medicine (TCM) demonstrate significant advantages in COPD treatment through multi-link, multi-pathway, and multi-mechanism intervention. Therefore, exploring the essence of COPD pathogenesis and discovering effective TCM treatment drugs through the application of TCM principles and prescriptions is a key focus of modern research. Animal models are of paramount importance in medical research. It is the first consideration to select appropriate animals, adopt reasonable modeling methods to replicate stable animal models that closely resemble the clinical manifestations and pathophysiological characteristics of COPD, and use appropriate evaluation methods to determine the success of COPD animal models in experimental research. The core of experimental research lies in observing the intervention effect of TCM on COPD animal models, exploring the specific pathways and regulatory mechanisms of TCM on COPD disease, and finding TCM monomers, single herbs, and TCM formulas with definite curative effects. At present, animal model research on COPD mainly involves model establishment, model evaluation, efficacy observation, mechanism exploration, and other aspects. In recent years, there has been no systematic organization, update, and reflection on the relevant research on TCM intervention in COPD animal models. This study reviewed the selection of animals for the COPD model, methods for establishing COPD animal models, model evaluation methods, and the intervention effects of TCM on COPD animal models. It aims to grasp the current research status and identify existing problems for further improvement, in order to provide evidence and support for scientific research and clinical treatment of COPD.
5.Yimei Baijiang Formula Treats Colitis-associated Colorectal Cancer in Mice via NF-κB Signaling Pathway
Qian WU ; Xin ZOU ; Chaoli JIANG ; Long ZHAO ; Hui CHEN ; Li LI ; Zhi LI ; Jianqin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):119-130
ObjectiveTo explore the effects of Yimei Baijiang formula (YMBJF) on colitis-associated colorectal cancer (CAC) and the nuclear factor kappaB (NF-κB) signaling pathway in mice. MethodsSixty male Balb/c mice of 4-6 weeks old were randomized into 6 groups: Normal, model, capecitabine (0.83 g
6.Animal Model of Chronic Obstructive Pulmonary Disease and Intervention Effect of Traditional Chinese Medicine: A Review
Jiyu ZOU ; Lijian PANG ; Tianjiao WANG ; Ningzi ZANG ; Zhongxue ZHAO ; Yongming LIU ; Qi SI ; Tianya CAO ; Xuenan MA ; Ying WANG ; Jiaran WANG ; Xiaodong LYU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):294-303
Chronic obstructive pulmonary disease (COPD), as one of the three major causes of death, is a complex systemic disease with high prevalence, high mortality, high disability, frequent acute exacerbations, and a variety of pulmonary complications. The pathogenesis is complex. Western medicine has no effective specificity scheme for a complete cure. However, multiple-component and multiple-target characteristics of traditional Chinese medicine (TCM) demonstrate significant advantages in COPD treatment through multi-link, multi-pathway, and multi-mechanism intervention. Therefore, exploring the essence of COPD pathogenesis and discovering effective TCM treatment drugs through the application of TCM principles and prescriptions is a key focus of modern research. Animal models are of paramount importance in medical research. It is the first consideration to select appropriate animals, adopt reasonable modeling methods to replicate stable animal models that closely resemble the clinical manifestations and pathophysiological characteristics of COPD, and use appropriate evaluation methods to determine the success of COPD animal models in experimental research. The core of experimental research lies in observing the intervention effect of TCM on COPD animal models, exploring the specific pathways and regulatory mechanisms of TCM on COPD disease, and finding TCM monomers, single herbs, and TCM formulas with definite curative effects. At present, animal model research on COPD mainly involves model establishment, model evaluation, efficacy observation, mechanism exploration, and other aspects. In recent years, there has been no systematic organization, update, and reflection on the relevant research on TCM intervention in COPD animal models. This study reviewed the selection of animals for the COPD model, methods for establishing COPD animal models, model evaluation methods, and the intervention effects of TCM on COPD animal models. It aims to grasp the current research status and identify existing problems for further improvement, in order to provide evidence and support for scientific research and clinical treatment of COPD.
7.Mechanism study of SIRT3 alleviating oxidative-stress injury in renal tubular cells by promoting mitochondrial biogenesis via regulating mitochondrial redox balance
Yaojun LIU ; Jun ZHOU ; Jing LIU ; Yunfei SHAN ; Huhai ZHANG ; Pan XIE ; Liying ZOU ; Lingyu RAN ; Huanping LONG ; Lunli XIANG ; Hong HUANG ; Hongwen ZHAO
Organ Transplantation 2026;17(1):86-94
Objective To elucidate the molecular mechanism of sirtuin-3 (SIRT3) in regulating mitochondrial biogenesis in human renal tubular epithelial cells. Methods Cells were stimulated with different concentrations of H2O2 and divided into four groups: control (NC), 50 μmol/L H2O2, 110 μmol/L H2O2 and 150 μmol/L H2O2. SIRT3 protein expression was then measured. SIRT3 was knocked down with siRNA, and cells were further assigned to five groups: control (NC), negative-control siRNA (NCsi), SIRT3-siRNA (siSIRT3), NCsi+H2O2, and siSIRT3+H2O2. After 24 h, cellular adenosine triphosphate (ATP) and mitochondrial superoxide anion (O2•−) levels were determined, together with mitochondrial expression of SIRT3, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), superoxide dismutase 2 (SOD2), acetylated-SOD2 and adenosine monophosphate activated protein kinase α1 (AMPKα1). Results The 110 and 150 μmol/L H2O2 decreased SIRT3 protein (both P<0.05). ATP and mitochondrial O2•− did not differ between NC and NCsi groups (both P>0.05). Compared to the NCsi group, the siSIRT3 group exhibited elevated O2•− level, decreased SIRT3 protein and increased expression levels of SOD2 and acetylated SOD2 protein (all P<0.05). Compared to the NCsi group, the NCsi+H2O2 group exhibited decreased cellular ATP levels, elevated mitochondrial O2•− levels, and reduced protein expression levels of SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 (all P<0.05). Compared with the siSIRT3 group, the siSIRT3+H2O2 group showed a decrease in cellular ATP levels, an increase in mitochondrial O2•− levels, a decrease in SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 protein expression levels and a decrease in acetylated SOD2 protein expression levels (all P<0.05). Compared with the NCsi+H2O2 group, the siSIRT3+H2O2 group showed a decrease in cellular ATP levels, an increase in mitochondrial O2•− levels, a decrease in SIRT3, AMPKα1, PGC-1α and NRF1, TFAM protein expression levels, and an increase in SOD2 and acetylated SOD2 protein expression levels (all P<0.05). Conclusions SIRT3 promotes mitochondrial biogenesis in tubular epithelial cells via the AMPK/PGC-1α/NRF1/TFAM axis, representing a key mechanism through which SIRT3 ameliorates oxidative stress-induced mitochondrial dysfunction.
8.Clinical Observation of Modified Huanglian Wendantang in Treatment of Cardiovascular Risk Factors in Patients with Metabolic Syndrome Under Guidance of Treating Disease before Its Onset
Yi HAN ; Yubo HAN ; Guoliang ZOU ; Ruinan WANG ; Chunli YAO ; Xinyu DONG ; Li LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):142-149
ObjectiveTo observe the clinical effect of modified Huanglian Wendantang on cardiovascular risk factors in patients with metabolic syndrome under the guidance of treating disease before its onset. MethodsA total of 82 patients with metabolic syndrome treated in the First Affiliated Hospital of Heilongjiang University of Chinese Medicine from July 2023 to July 2024 were selected and allocated into an observation group (41 cases) and a control group (41 cases) by the random number table method. The control group received routine treatment, and the observation group was treated with modified Huanglian Wendantang on the basis of routine treatment. Both groups were treated for 8 weeks. The therapeutic effects on TCM symptoms after treatment in the two groups were evaluated. The levels of obesity degree indicators, blood pressure indicators, glucose and lipid metabolism indicators, inflammatory factors, and vascular endothelial function indicators before and after treatment in the two groups were measured, and the treatment safety was evaluated. ResultsAfter treatment, the total response rate of TCM symptoms in the observation group was 97.56% (40/41), which was higher than that (87.80%, 36/41) in the control group (χ2=5.205, P<0.05). After treatment, both groups showed declines (P<0.05) in systolic blood pressure (SBD), diastolic blood pressure (DBP), triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), fasting blood glucose, 2-hour postprandial blood glucose (2 h PG), glycosylated hemoglobin (HbA1c), fasting insulin (FINS), Homeostatic Model Assessment for Insulin Resistance (HOMA-IR), body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), leptin (LEP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), endothelin-1 (ET-1), and inducible nitric oxide synthase (iNOS). Moreover, the declines in the observation group were more obvious than those in the control group (P<0.05, P<0.01). After treatment, both groups showed elevated levels of high density lipoprotein cholesterol (HDL-C), adiponectin (ADP), nitric oxide (NO), and endothelial nitric oxide synthase (eNOS) (P<0.05), and the above indexes in the observation group were higher than those in the control group (P<0.01). ConclusionUnder the guidance of the thought of treating disease before its onset, modified Huanglian Wendantang was used to treat patients with metabolic syndrome. The decoction improved the clinical efficacy by ameliorating IR to improve insulin sensitivity, reducing inflammation, and protecting the vascular endothelial function. It inhibits cardiovascular risk factors without inducing adverse reactions, being worthy of clinical application and promotion.
9.Surveillance results of respiratory syncytial virus outbreaks in kindergarten and school in Shenzhen, 2017-2023
WANG Xin, FANG Shisong, WU Weihua, LIU Hui, SUN Ying, ZOU Xuan, TANG Xiujuan
Chinese Journal of School Health 2025;46(3):435-437
Objective:
To analyze respiratory syncytial virus(RSV) outbreaks surveillance results and the epidemiological characteristics in kindergarten and school in Shenzhen during 2017-2023 , so as to provide a scientific reference for control and prevention of RSV.
Methods:
Epidemiological data and surveillance results of RSV outbreaks in kindergarten and school from 2017 to 2023 were collected for descriptive analyses.
Results:
A total of 31 RSV outbreaks were identified in kindergarten and school in 2017-2023 in Shenzhen, 346 cases were reported, the average incidence rate was 22.02%. The most annual RSV outbreaks were reported in 2020 with 14 outbreaks, followed by 8 outbreaks in 2023. A total of 64.52% of RSV outbreaks were identified in kindergarten with rest occurring in primary school or middle school. The greatest monthly count of outbreak was 18 (58.06%) in September, followed by 3 outbreaks (9.68%) in March and October. A total of 244 swab samples were collected, 169 samples were positive for respiratory viruses, the positive rate was 69.26%, 121 samples were positive for RSV,from 31 respiratory syncytical virus outbreaks 57 and samples were positive for other respiratory viruses(9 samples were positive for two respiratory viruses). A toral of 14(45.16%) outbreaks are caused by RSV alone, 17 outbreaks (54.84%) were caused by RSV and other respiratory viruses.
Conclusions
Most RSV outbreaks in kindergarten and school are reported after 2020 in Shenzhen, most RSV outbreaks occur in kindergarten, peak seasons of RSV outbreaks are autumn and spring.
10.Causal relationship between mental disorders and hypercholesterolemia: a Mendelian randomization analysis
Qian ZOU ; Ni TANG ; Huanhui LIU ; Hanjing ZHANG ; Xiaojie MA
Sichuan Mental Health 2025;38(2):123-130
BackgroundMetabolic syndrome (MetS) is highly prevalent in patients with mental disorders, including elevated diastolic or systolic blood pressure, elevated fasting glucose, hypercholesterolemia, abdominal obesity and so on. As an important component of MetS, the relationship between hypercholesterolemia and mental disorder has been extensively reported, whereas few genome-wide association studies (GWAS) have been conducted to identify the causal role of mental disorders in hypercholesterolemia. ObjectiveTo explore the potential causal relationship between mental disorders and hypercholesterolemia by two-sample Mendelian randomization (MR) method. MethodsSummary data from GWAS were analyzed. Single nucleotide polymorphisms (SNPs) strongly associated with mental disorders were chosen as instrumental variables, and hypercholesterolemia was used as outcome variable. MR analysis utilized inverse-variance weighted (IVW), MR-Egger regression and weighted median estimation (WME) as the primary analytical tool, and supplemented by simple mode (SM) and weighted mode (WM). The causal relationship between mental disorders and the risk of hypercholesterolemia was illustrated in terms of odds ratio (OR). ResultsA total of 36 SNPs associated with mental disorders were identified as instrumental variables. The primary findings from IVW revealed existence of a causal relationship between mental disorders and hypercholesterolemia (IVW: OR=1.067, 95% CI: 1.026~1.109, P=0.001). Findings from the additional methods (MR-Egger regression, WME, SM, WM) were basically consistent with those reported in IVW method. Further verification indicated that the causal relationship between mental disorders and the risk of hypercholesterolemia was not affected by genetic polymorphism (P>0.05). The absence of heterogeneity was confirmed through Cochran's Q test and MR-Egger regression (P>0.05). Furthermore, no causal association in the reverse direction was found (P>0.05). ConclusionThere is a causal relationship between mental disorders and hypercholesterolemia, and patients with mental disorders may have an increased probability of suffering from hypercholesterolemia.


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