1.RUNX3 regulates FAP to influence the proliferation of mouse lung primary fibroblasts
Junbo YOU ; Xianchen WANG ; Hui LING ; Jiahao FAN ; Qi CHEN ; Hui TAO ; Jiming SHA
Acta Universitatis Medicinalis Anhui 2026;61(4):606-611
ObjectiveTo investigate the role of runt-related transcription factor 3 (RUNX3) in transforming growth factor-β1 (TGF-β1)-induced activation of mouse primary pulmonary fibroblasts (PFs), and its effects on fibroblast activation protein (FAP) expression, cell proliferation, and collagen synthesis. MethodsPFs were isolated from C57BL/6 mice and cultured. A RUNX3 knockdown model was established using small interfering RNA (siRNA). Cells were assigned to the control group (Control), TGF-β1-treated group (TGF-β1), negative control group (TGF-β1+siRNA-NC), and RUNX3-silenced group (TGF-β1+si-RUNX3). In addition, a RUNX3 overexpression rescue experiment was performed based on TGF-β1 stimulation. Protein and mRNA levels of RUNX3, FAP, and typeⅠcollagen (COL1A1) were measured by Western blot and reverse transcription quantitative real-time PCR (RT-qPCR). Cell proliferation was assessed using CCK-8 and EdU assays. Co-expression of COL1A1 and FAP was examined by double immunofluorescence staining. ResultsCompared with the Control group, RUNX3, FAP, and COL1A1 expression levels were upregulated in PFs in the TGF-β1 group (P<0.01). The CCK-8 assay showed that the absorbance value was reduced in the RUNX3 knockdown group compared with the negative control group (P<0.01). Consistently, the EdU assay demonstrated a lower proportion of EdU-positive cells in the RUNX3 knockdown group than in the negative control group (P<0.01). Immunofluorescence double staining revealed decreased fluorescence intensities of COL1A1 and FAP in the RUNX3 knockdown group relative to the negative control. Under RUNX3 overexpression conditions, these fluorescence signals exhibited a partial rebound (P<0.01). ConclusionRUNX3 in TGF-β1-induced PFs may promote cell proliferation and collagen synthesis by positively regulating FAP expression. Targeting the RUNX3/FAP axis may represent a potential therapeutic strategy for pulmonary fibrosis.
2.Effect of RUNX3 on the activation, proliferation, and migration capabilities of hepatic stellate cells
Hui LING ; Xianchen WANG ; Junbo YOU ; Jiahao FAN ; Xiao CUI ; Jiming SHA ; Liquan YU
Acta Universitatis Medicinalis Anhui 2026;61(2):277-284
ObjectiveTo investigate the effects of targeted silencing of Runt-related Transcription Factor 3 (RUNX3) on the proliferation and migration of Mouse Hepatic Stellate Cells (HSCs), as well as subsequent collagen deposition. MethodsMouse hepatic stellate cell line (JS-1) was selected and then morphologically observed and identified under a microscope. After the cells had fully adhered, they were treated with 5 ng/mL of transforming growth factor beta 1 (TGF-β1) for 24 hours to induce hepatic stellate cell activation. Furthermore, a RUNX3 silencing model was established using RUNX3 lentiviral infection. The experiment was divided into four groups: Control group, TGF-β1 group, TGF-β1+siRNA-NC group, and TGF-β1+siRNA-RUNX3 group. Protein expression changes of RUNX3, alpha-smooth muscle actin (α-SMA), and Alpha 1 type I collagen (Collagen I) were detected using Western blot method. Cellular immunofluorescence assays were employed to investigate the deposition changes of α-SMA and RUNX3 in hepatic stellate cells. RT-qPCR was utilized to examine the mRNA expression changes of RUNX3, α-SMA, and Collagen I. The proliferative capacity of hepatic stellate cells was assessed using Edu staining. The migratory ability of hepatic stellate cells was evaluated through wound healing assays and Transwell migration experiments. ResultsCompared with Control group, a significant elevation in RUNX3 was observed in the TGF-β1-induced activated HSCs (P<0.01). Meanwhile, the protein and mRNA levels of fibrosis-related markers and α-SMA and Collagen I were significantly upregulated (P<0.001). Additionally, the proliferation and migration capabilities of HSCs were significantly enhanced (P<0.001). In contrast, when compared to TGF-β1+siRNA-NC group, TGF-β1+siRNA-RUNX3 group exhibited a notable decrease in RUNX3 and other related indicators, such as the protein and mRNA levels of α-SMA and Collagen I (P<0.05). Concurrently, the proliferation and migration capabilities of HSCs were significantly inhibited in TGF-β1+siRNA-RUNX3 group (P<0.01). ConclusionSilencing RUNX3 can inhibit the deposition of collagen and the proliferation and migration of hepatic stellate cells. Conversely, RUNX3 promotes the proliferation and migration capabilities of HSCs, thereby facilitating the activation of HSC.
3.Effect of targeted silencing of DNMT3A on collagen deposition, proliferation and migration activity of mouse lung fibroblasts
Xianchen Wang ; Junbo You ; Hui Ling ; Jiahao Fan ; Qi Chen ; Hui Tao ; Jiming Sha
Acta Universitatis Medicinalis Anhui 2025;60(1):66-72
Objective:
To investigate the effect of targeted silencing of DNA methyltransferase 3A(DNMT3A) on collagen deposition, proliferation and migration activity of mouse lung fibroblasts(PFs).
Methods:
In order to ensure the proliferation and migration activity of primary fibroblasts, the lung tissues of neonatal C57 suckling mice were taken, PFs were extracted after being sheared, and the morphology was observed and identified under the microscope. PFs cells were activated by 5 ng/ml TGF-β1for 24 h after cell attachment, and DNMT3A silencing model was constructed by small interfering RNA; The experiment was divided into control group, TGF-β1group, TGF-β1+ siRNA-NC group and TGF-β1+ siRNA-DNMT3A group. The protein expressions of DNMT3A, α-smooth muscle actin(α-SMA) and Collagen Ⅰ were detected by Western blot; Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression changes ofDNMT3A,α-SMAandCollagenⅠ. The proliferation ability of PFs was detected by CCK-8 and EdU staining; the migration ability of PFs was detected by scratch test and Transwell migration test.
Results:
Compared with the control group, TGF-β1induced the increase of DNMT3A in the activated PFs cell group(P<0.01), the protein and mRNA levels of fibrosis and proliferation related indicators α-SMA and Collagen Ⅰ also increased(allP<0.05), and the proliferation and migration ability of PFs increased(allP<0.000 1). Compared with the siRNA-NC group, the protein expression levels of DNMT3A(P<0.000 1) and related indicators α-SMA(P<0.01) and Collagen Ⅰ(P<0.01) significantly decreased in the DNMT3A silencing group by Western blot, and the mRNA levels ofDNMT3A,α-SMAandCollagenⅠby RT-qPCR also decreased(allP<0.001), and the proliferation(P<0.01) and migration ability(P<0.05) of PFs cells decreased compared with the control group.
Conclusion
Silencing DNMT3A can inhibit the deposition of collagen and the proliferation of PFs. DNMT3A can promote the proliferation and migration of PFs, and then promote the activation of PFs and the development of pulmonary fibrosis. This process may be regulated by DNA methylation modification.
4.Expert Consensus on the Ethical Requirements for Generative AI-Assisted Academic Writing
You-Quan BU ; Yong-Fu CAO ; Zeng-Yi CHANG ; Hong-Yu CHEN ; Xiao-Wei CHEN ; Yuan-Yuan CHEN ; Zhu-Cheng CHEN ; Rui DENG ; Jie DING ; Zhong-Kai FAN ; Guo-Quan GAO ; Xu GAO ; Lan HU ; Xiao-Qing HU ; Hong-Ti JIA ; Ying KONG ; En-Min LI ; Ling LI ; Yu-Hua LI ; Jun-Rong LIU ; Zhi-Qiang LIU ; Ya-Ping LUO ; Xue-Mei LV ; Yan-Xi PEI ; Xiao-Zhong PENG ; Qi-Qun TANG ; You WAN ; Yong WANG ; Ming-Xu WANG ; Xian WANG ; Guang-Kuan XIE ; Jun XIE ; Xiao-Hua YAN ; Mei YIN ; Zhong-Shan YU ; Chun-Yan ZHOU ; Rui-Fang ZHU
Chinese Journal of Biochemistry and Molecular Biology 2025;41(6):826-832
With the rapid development of generative artificial intelligence(GAI)technologies,their widespread application in academic research and writing is continuously expanding the boundaries of sci-entific inquiry.However,this trend has also raised a series of ethical and regulatory challenges,inclu-ding issues related to authorship,content authenticity,citation accuracy,and accountability.In light of the growing involvement of AI in generating academic content,establishing an open,controllable,and trustworthy ethical governance framework has become a key task for safeguarding research integrity and maintaining trust within the academic community.This expert consensus outlines ethical requirements across key stages of AI-assisted academic writing-including topic selection,data management,citation practices,and authorship attribution.It aims to clarify the boundaries and ethical obligations surrounding AI use in academic writing,ensuring that technological tools enhance efficiency without compromising in-tegrity.The goal is to provide guidance and institutional support for building a responsible and sustainable research ecosystem.
5.Ameliorative effects of sweet potato leaf extract on mammary gland oxidative stress and rumenmicrobiota in dairy goats under high concentrate feeding pattern
Ziqing XIU ; Ling ZHANG ; Junqiu ZHANG ; Yu CHEN ; Mgeni MUSA ; Yongjiang WU ; Juncai CHEN ; Yawang SUN ; You YANG
Chinese Journal of Veterinary Science 2025;45(9):1952-1964
This study aimed to investigate the effects of sweet potato leaf extract on production per-formance,systematic and mammary gland oxidative stress status and rumen microbiota of dairy goats fed high concentrate diets.Twenty Guanzhong dairy goats with same parity,similar lactation period(120±15)d and healthy body condition were selected and randomly divided into four groups:low-concentrate(LC),low-concentrate supplemented with 1%sweet potato leaf extract(LCS),high-concentrate(HC)and high-concentrate supplemented with 1%sweet potato leaf ex-tract(HCS).The experimental period was 35 days.The results showed that in the third week,milk yield in the HCS group was significantly higher than that in the LC and LCS groups(P<0.05).The content of lipopolysaccharide in the rumen fluid of the HC group was significantly higher than that of the other three groups(P<0.05),the content of malondialdehyde in the serum of the HC group was significantly higher than that of the LCS group(P<0.05),the content of reactive oxygen species,protein carbonyl,8-hydroxydeoxyguanosine in the milk of the HC group was sig-nificantly higher than that of the LC and LCS groups(P<0.05),GSH-Px in HCS group was sig-nificantly higher than that in the other three groups(P<0.05).After the addition of sweet potato leaf extract,there was an increasing trend in the content of Anabaena phylum at the phylum level.In the joint analysis of genera,rumen fluid LPS showed highly significant negative correlation with Succiniclasticum(P<0.01)and negative correlation with Prevotella(P<0.05).Valeric acid was negatively correlated with Prevotella(P<0.05).The pH value was negatively correlated with Treponema(P<0.05).Butyric acid was positively correlated with Anaeroplasma(P<0.05).In conclusion,the addition of sweet potato leaf extract to the diet can increase milk production and al-leviate the state of mammary gland oxidative stress,as well as improving rumen microbial diversi-ty of dairy goats.
6.Invasive pulmonary aspergillosis assisted in clinical diagnosis by chest CT pulmonary angiography: three cases report
Dengmei TIAN ; Jianhua YOU ; Jiong HU ; Ling WANG
Chinese Journal of Hematology 2025;46(10):972-976
Chest CT pulmonary angiography (CTPA) has certain auxiliary diagnostic value for the clinical diagnosis of invasive pulmonary aspergillosis (IPA) . Three patients with hematological malignancies were reported, including 2 ones after allogeneic hematopoietic stem cell transplantation and 1 ones after chemotherapy for refractory recurrent leukemia. Each patient was treated with antibiotics for at least 48 hours after the onset of fever, they all underwent chest high-resolution CT (HRCT) scans without fever resolution. CT revealed at least one dense pulmonary consolidation shadow with a diameter greater than 10 mm, and subsequently a CTPA examination was performed to observe the effect of CTPA imaging signs for the diagnosis of IPA. There were 2 patients with positive vascular occlusion sign (VOS) and 1 patient with negative VOS detected by CTPA. Among them, 2 patients with positive VOS were diagnosed with possible IPA and received with diagnosis-driven antifungal treatment, which improved their conditions. One patient with negative VOS sign was diagnosed with diffuse large B-cell lymphoma involving the lungs. After receiving anti-lymphoma treatment, the lesions significantly reduced in size. The vascular occlusion sign detected by CTPA is relatively characteristic. For high-risk IPA patients, it helps to improve the specificity of imaging diagnosis and guide clinical treatment decisions.
7.Engineering CHO Cell Lines to Stably Express B4GALT1,ST6GAL1,and GnTⅢ with Site-directed Integration
Xian-Hong LI ; Run-Qing JIA ; You-Liang WANG ; Wei-Ling MAN ; Tian-Hao ZHU ; Xin-Long YAN ; Yan-Li LIN
Chinese Journal of Biochemistry and Molecular Biology 2025;41(4):576-585
Glycoengineering was carried out in the mammalian cell line CHO for the production of pro-tein-based drugs.Firstly,the genome sequence of the Rosa26 locus of CHO cells was determined,the gRNA sequences were designed,and the landing pad was integrated into the Rosa26 locus of CHO cells by CRISPR/Cas9 technology.Three targeting vectors co-expressed by glycosyltransferases,which are β-1,4 galactosyltransferase(B4GALT1),α-2,6-sialyltransferase 1(ST6GAL1)and N-acetaminoglycosyl-transferase Ⅲ(GnT Ⅲ),were constructed by overlapping PCR and seamless ligation technology,and the three glycosyltransferase genes were integrated into the CHO Rosa26 locus by Cre enzyme-mediated cassette exchange technology.PCR confirmed that three glycosyltransferases had been successfully site-directed integrated into the Rosa26 site.The mRNA expression levels of the three glycosyltransferases were more than 50 000-fold by qRT-PCR,and the protein expression levels of the three glycosyltrans-ferases were more than 4-fold via western blotting(P<0.001).A CHO-engineered cell line with three glycosyltransferases integrated into Rosa26 site was successfully constructed.
8.Invasive pulmonary aspergillosis assisted in clinical diagnosis by chest CT pulmonary angiography: three cases report
Dengmei TIAN ; Jianhua YOU ; Jiong HU ; Ling WANG
Chinese Journal of Hematology 2025;46(10):972-976
Chest CT pulmonary angiography (CTPA) has certain auxiliary diagnostic value for the clinical diagnosis of invasive pulmonary aspergillosis (IPA) . Three patients with hematological malignancies were reported, including 2 ones after allogeneic hematopoietic stem cell transplantation and 1 ones after chemotherapy for refractory recurrent leukemia. Each patient was treated with antibiotics for at least 48 hours after the onset of fever, they all underwent chest high-resolution CT (HRCT) scans without fever resolution. CT revealed at least one dense pulmonary consolidation shadow with a diameter greater than 10 mm, and subsequently a CTPA examination was performed to observe the effect of CTPA imaging signs for the diagnosis of IPA. There were 2 patients with positive vascular occlusion sign (VOS) and 1 patient with negative VOS detected by CTPA. Among them, 2 patients with positive VOS were diagnosed with possible IPA and received with diagnosis-driven antifungal treatment, which improved their conditions. One patient with negative VOS sign was diagnosed with diffuse large B-cell lymphoma involving the lungs. After receiving anti-lymphoma treatment, the lesions significantly reduced in size. The vascular occlusion sign detected by CTPA is relatively characteristic. For high-risk IPA patients, it helps to improve the specificity of imaging diagnosis and guide clinical treatment decisions.
9.A new trend in the development of laboratory medicine:clinical laboratory omics
Yingqiang DANG ; Yao JIANG ; Qian WU ; Ling MENG ; Chongge YOU
International Journal of Laboratory Medicine 2025;46(15):1873-1878
With the rapid development of artificial intelligence and machine learning algorithms in the med-ical field,as well as the high-quality development requirements put forward by the state for hospitals and vari-ous departments,the development of laboratory medicine shows a new trend.The development of clinical labo-ratory has evolved from the initial test suite to the test pathway based on clinical pathway and diagnosis-relat-ed groups payment,and then to clinical laboratory omics.Clinical laboratory omics refers to the use of high-throughput methods to obtain a large number of laboratory project results data.Combined with the clinical characteristics of patients,statistical methods and machine learning algorithms are commonly used to reveal the information behind a large number of medical data to assist clinicians in diagnosis and treatment.Clinical laboratory omics may be a new trend in the future development of laboratory medicine,and it is likely to play an important role in the field of laboratory medicine.
10.Engineering CHO Cell Lines to Stably Express B4GALT1,ST6GAL1,and GnTⅢ with Site-directed Integration
Xian-Hong LI ; Run-Qing JIA ; You-Liang WANG ; Wei-Ling MAN ; Tian-Hao ZHU ; Xin-Long YAN ; Yan-Li LIN
Chinese Journal of Biochemistry and Molecular Biology 2025;41(4):576-585
Glycoengineering was carried out in the mammalian cell line CHO for the production of pro-tein-based drugs.Firstly,the genome sequence of the Rosa26 locus of CHO cells was determined,the gRNA sequences were designed,and the landing pad was integrated into the Rosa26 locus of CHO cells by CRISPR/Cas9 technology.Three targeting vectors co-expressed by glycosyltransferases,which are β-1,4 galactosyltransferase(B4GALT1),α-2,6-sialyltransferase 1(ST6GAL1)and N-acetaminoglycosyl-transferase Ⅲ(GnT Ⅲ),were constructed by overlapping PCR and seamless ligation technology,and the three glycosyltransferase genes were integrated into the CHO Rosa26 locus by Cre enzyme-mediated cassette exchange technology.PCR confirmed that three glycosyltransferases had been successfully site-directed integrated into the Rosa26 site.The mRNA expression levels of the three glycosyltransferases were more than 50 000-fold by qRT-PCR,and the protein expression levels of the three glycosyltrans-ferases were more than 4-fold via western blotting(P<0.001).A CHO-engineered cell line with three glycosyltransferases integrated into Rosa26 site was successfully constructed.


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