Acta Universitatis Medicinalis Anhui

Journal Information

Superintend by

Anhui Provincial Education Department

Sponsored by

Anhui Medical University

Editor-in-chief

Weng Jianping

Edited and Published by

Editorial Board of Acta Universitatis Medi-cinalis Anhui Meishan Road , Hefei 230032

Printed by

Anhui Lianzhong Printing Limited

Company

Domestic Subscription

Local Post Office , China

Domestic Distribution

Hefei Post Office of Anhui Province, China

Post Subscription Number

26 - 36

Foreign Distribution

China International Book Trading

Corporation

No.35, chegongzhuang West Road, Haidian District, Beijing, 100048,

China Code No . MO5933

Price

￥:10.00/Issue, 120.00/Year

CSSN

ISSN 1000 - 1492

CN 34 - 1065/R

Tel

(0551)65161103, 65161192

E-mail

aydxb@vip.163.com (Submission)

aydxbsg@163.com (Review)

CODEN

AYDXAG

Web Address

https://www.aydxb.cn/

https://yike.cbpt.cnki.net/portal

Official Account

WeChat applet

Display method:

Csf1r-Cre-mediated yellow fluorescent protein can effectively label tissue-resident macrophages in various tissues

Li Xiaoyu1, Zhao Dianyuan2,3, Zhang Shengquan1, Yang Jing1,2,3, Tang Li1,2,3

DOI:

PDF(3) browse(99)

after spinal cord injury through PI3K/AKT/NF-κB signaling pathway

Ye Lei, You Tao

DOI:

PDF(8) browse(91)

abstract:
To explore the mechanism of hesperidin on hindlimb motor function in mice with spinal cord injury (SCI). Methods Oxygen glucose deprivation (OGD) was used to induce SCI in PC12 neuronal cells. CCK-8 assay determined the optimal hesperidin concentration (0, 10, 20, 30, 40, 50, 60 mmol/L) for subsequent experiments after 24h treatment. PC12 cells were divided into Control group, OGD group and Hesperidin (30 mmol/L)+OGD groups. Hoechst staining assessed apoptosis; Western blot (WB), qPCR and immunofluorescence (IF) detected the expression of iNOS, CD206 and TNF-α . Flow cytometry measured apoptosis rates, and WB examined the impact of hesperidin on PI3K, p-PI3K, AKT, p-AKT, and NF-κB expression. Forty-five healthy male C57BL/6 mice were randomly divided into Sham group, Sci group and Hesperidin groups. Sham and Sci groups received 0.1 mL PBS, while the Hesperidin group received 0.1 mL hesperidin. After modeling, iNOS, CD206, and TNF-α expression levels, spinal cord cavitation, and hindlimb motor function were evaluated. Results Hesperidin at 10–30 mmol/L for 24h did not significantly affect PC12 cell activity. Hoechst staining showed reduced apoptosis in PC12 cells after 24h of 30 mmol/L hesperidin treatment. Compared to the OGD group, Hesperidin+OGD group had decreased mRNA and protein expression of inﬂammatory factors (iNOS and TNF-α) and increased expression of the anti-inﬂammatory factor (CD206). Hesperidin treatment inhibited PI3K/AKT/NF-κB pathway activation, with similar results in mouse experiments. Compared to Sci group, Hesperidin group had reduced spinal cord cavitation area and improved hindlimb motor function. Conclusion Hesperidin may improve hindlimb motor function in mice with SCI by downregulating the phosphorylation level of the PI3K/AKT/NF-κB pathway, inhibiting inﬂammatory factor release, promoting anti-inﬂammatory factor release, regulating the inﬂammatory microenvironment after SCI, and suppressing the acute inﬂammatory response.

Impacts of remifentanil on proliferation, apoptosis and Wnt/β-catenin pathway of breast cancer MDA-MB-231 cells

Zhang Yang1, Liu Wenwen2, Miao Huanhuan3, Yang Guang1

DOI:

PDF(0) browse(80)

abstract:
To investigate the impacts of remifentanil on the proliferation, apoptosis and Wnt/β-catenin pathway of breast cancer MDA-MB-231 cells. Methods Different concentrations of remifentanil (0, 0.5, 2.5, 5.0, 10.0, 20.0, 40.0 µg/mL) were used to treat human normal breast MCF-10A cells and breast cancer MDA-MB-231 cells to screen for experimental concentrations of remifentanil. MDA-MB-231 cells were divided into control group, remifentanil low (5.0 µg/mL), medium (10.0 µg/mL), and high (20.0 µg/mL) concentration groups, and remifentanil+SKL2001 (Wnt/β-catenin pathway agonist) group. MTT assay and colony formation assay were applied to detect cell proliferation ability. Flow cytometry was applied to detect cell apoptosis and cell cycle changes. Western blot was applied to detect protein expression related to cell proliferation, apoptosis, and the Wnt/β-catenin signaling pathway. Results Remifentanil at concentrations of 5.0, 10.0, and 20.0 µg/mL could reduce the viability of MDA-MB-231 cells and had no prominent toxicity to MCF-10A cells. Compared with the control group, the optical density value of cell proliferation, colony formation number, proportion of S-phase cells, and the protein levels of c-Myc, Cyclin D1, B lymphoblastoma 2 (Bcl-2), precursor of cysteine aspartate protease-3 (pro-caspase-3) and β-catenin were lower in the low, medium, and high concentration remifentanil groups (P<0.05), while the proportion of G0/G1 phase cells, apoptosis rate, early apoptosis rate, the protein levels of Bcl-2 associated X protein (Bax) and cleaved cysteine aspartate protease-3 (Cleaved caspase-3), and glycogen synthase kinase-3β (GSK-3β) were higher (P<0.05). SKL2001 could weaken the effects of remifentanil on the proliferation and apoptosis of MDA-MB-231 cells. Conclusion Remifentanil inhibits the proliferation of MDA-MB-231 cells, induces apoptosis and cell cycle arrest, and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway activation.

BMP-2 gene-modified bone marrow mesenchymal stem cells combined with nano-hydroxyapatite/polyamide 66 implants improve atrophic nonunion of bone in rats

Huang Yong, Ma Jianwen, Li Yu, Jing Qingling, Zhang Qin, Li Changshuai

DOI:

PDF(2) browse(73)

abstract:
（Dept of Trauma Orthopedics, Affiliated Hospital of Qinghai University Xining 810001）Abstract Objective To investigate the therapeutic effect of bone marrow mesenchymal stem cells (BMSCs) modified by bone morphogenetic protein (BMP)-2 gene combined with nano-hydroxyapatite (nHA)/polyamide 66 (PA66) on femoral nonunion in rats. Methods Rat BMSCs were isolated and divided into the stent+BMSCs group, stent+BMSCs+rhBMP-2 group,and stent+BMSCs+Ad-BMP-2 group; human recombinant BMP-2 (rhBMP-2) or adenovirus-infected BMP-2 (Ad-BMP-2) vector was transfected into BMSCs and loaded onto nHA/PA66 stent materials. Flow cytometry was used to detect that BMSCs expressed specific surface markers. Cell proliferation was detected by MTT assay, related bioactive factors [platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), osteocalcin (OCN), osteonectin (ON)] were detected by ELISA, alkaline phosphatase (ALP) activity was detected, and cell growth and adhesion were observed by scanning electron microscopy (SEM). In the atrophic nonunion model of SD rats, the stent+BMSCs+rhBMP-2 or stent+BMSCs+Ad-BMP-2 complex was implanted into the defect area, which was divided into the rhBMP-2 group and the Ad-BMP-2 group, and the therapeutic effect was evaluated by X-ray and Micro CT. Results The surface of the nHA/PA66 stent was smooth and porous, and BMSCs adhered well. Flow cytometry showed high expression of CD29 and CD90 and low expression of CD45 and CD34 in BMSCs. MTT showed that the cells proliferated rapidly after 72 h. Compared with the stent+BMSCs group and the stent+BMSCs+rhBMP-2 group, the ALP activity, the expressions of PDGF, TGF-β, VEGF,FGF, OCN, and ON in the stent+BMSCs+Ad-BMP-2 group were significantly up-regulated (P<0.05). 12 weeks after surgery, the stent complex in the Ad-BMP-2 group of rats was completely wrapped by newly formed cortical bone, and the surface was smooth and intact. X-ray showed that the complex in the Ad-BMP-2 group was completely wrapped by newly formed cortical bone, and the recovery degree was better than that in the rhBMP-2 group. Micro CT results showed that compared with the rhBMP-2 group, the bone volume fraction, trabecular thickness, trabecular number, bone mineral density, and bone mineral content in the Ad-BMP-2 group all increased 12 weeks after surgery (P<0.05), and the trabecular separation level decreased (P<0.05). Conclusion Ad-BMP-2-transfected BMSCs combined with nHA/PA66 stent material can express bioactivity more effectively, provide a continuous and good microenvironment for the proliferation and differentiation of BMSCs, which is beneficial to promoting local osteogenic activity and bone defect repair.

Preliminary exploration of the value of combined detection of folate receptor-positive circulating tumor cells and hemostatic function in improving metastasis diagnosis of gastrointestinal tumor

Xiao Yanlin1, Ji Duxin2, Feng Qingqing1, He Huidan1, Bian Maohong1

DOI:

PDF(1) browse(72)

abstract:
To investigate the diagnostic value of combined detection of folate receptor-positive circulating tumor cells (FR+-CTCs) and hemostatic function indicators in improving the diagnosis of gastrointestinal tumors (GITs) metastasis. Methods A retrospective analysis was conducted on the clinical data of 186 patients aged 18 to 80 years who were diagnosed with gastrointestinal tumors via pathology and received treatment, including data on FR+-CTCs, hemostatic function indicators, and pathological staging. The collected data encompassed FR+-CTCs levels, coagulation parameters, and pathological staging. Statistical analysis included t-tests, chi-square tests, fisher’s exact test, logistic regression analysis, and receiver operating characteristic (ROC) curves to assess the diagnostic value of combined FR+-CTCs and coagulation parameters in detecting tumor metastasis. Results FR+-CTCs levels and positive rates demonstrated significant associations with clinicopathological characteristics (gender, histological type, N staging) in GITs patients (P<0.05). In patients with metastasis, elevated fibrinogen levels were observed, while both thrombin time and platelet counts showed significant increases in N1 ~ N3 stages (P<0.05). Logistic regression analysis showed that PLT and antithrombin III (AT-III) were independent risk factors for GITs metastasis (P<0.05). The areas under the ROC curves for predicting GITs metastasis were 0.678 (95%CI: 0.540-0.816) and 0.664 (95%CI: 0.512~0.815), respectively. When combining multiple factors, including FR⁺-CTCs, PLT, AT-III, pathological type, FIB, TT, and gender, for the diagnosis of GITs metastasis, the AUC increased to 0.757 (95%CI: 0.621-0.893), indicating higher sensitivity and specificity compared to using each indicator alone. Conclusion The combined detection of FR+-CTCs and anticoagulation function indicators has a higher diagnostic value for the diagnosis of GITs, providing a valuable basis for the early diagnosis of GITs, especially in metastasis surveillance.

Establishment and identification of hepatocyte-specific Tmem121 knockout mouse model

Wang Yue1, He Guoliang1, Li Lanyu2, Wu Qian3, Zhou Junmei1,4,5,6

DOI:

PDF(0) browse(63)

Expression and optimization of recombinant human coagulation factor X in HEK293 cells

Feng Jianing1, Zou Sen2, Zhao Zelin1, Li Xiaoxiao1, Zhang Zhifei1, Yang Zhaoyong2

DOI:

PDF(1) browse(60)

abstract:
To perform transient transfection of recombinant human Factor X (rhFX) into HEK293 cells, to optimize transfection parameters, to develop a high-yield in vitro expression system for rhFX production, and to assess the biological activity of expressed rhFX. Methods The eukaryotic expression vector pcDNA3.1-EGFP-FX was constructed by inserting the FX gene into pcDNA3.1-EGFP and subsequently transfected into HEK293 cells to validate the transfection system efficiency. The recombinant expression vector pcDNA3.1-FX was generated through ligation of the FX gene fragment with pcDNA3.1, followed by liposome-mediated transfection into HEK293 cells. The expression of rhFX in the cell supernatant was analyzed by Western blot and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Transfection conditions were systematically optimized, and rhFX concentration was quantified by enzyme-linked immunosorbent assay (ELISA). The coagulation bioactivity of rhFX was evaluated through prothrombin time (PT) assay and chromogenic substrate method. Results Recombinant human Factor X (rhFX) was successfully expressed in the supernatant ofHEK293 cells. Optimal expression was achieved at 80% cell confluence with a ratio of plasmid DNA to transfection reagent as 1:2. Peak rhFX production was observed on day 3 post-transfection. Triplicate ELISA measurements demonstrated a maximum rhFX concentration of 5.20 ng/mL in the supernatant. The prothrombin time (PT) of rhFX-containing supernatant was significantly shorter (P < 0.0001) compared to control supernatant from pcDNA3.1 empty vector-transfected cells. The half maximal inhibitory concentration (IC50) of etoxaban was determined to be 1.449 nmol/L using the chromogenic substrate method based on rhFX, which was in the same order of magnitude as the reported 0.78 nmol/L in the literature. Conclusion HEK293 cells successfully express biologically active recombinant human Factor X (rhFX) protein, laying a foundation for advancing the development of rhFX-based therapeutics.

Construction and evaluation of a prognostic nomogram prediction model for patients with coronary heart disease based on Lp-PLA2 、LP(a), and clinical risk factors

Wang Tianqi, Hu Zeping, Zhu Xuetao

DOI:

PDF(3) browse(69)

abstract:
To construct and to validate a nomogram prediction model based on Lipoprotein-associated phospholipase A2 (Lp-PLA2) and Lipoprotein(a) [LP(a)] for predicting the risk of major adverse cardiovascular events (MACE) in patients with coronary heart disease (CHD). Methods A retrospective analysis was conducted on the clinical data of 442 patients with coronary heart disease (CHD). Among them, 411 patients who completed follow-up were randomly divided into a training set (288 cases) and a validation set (123 cases) at a 7:3 ratio. Independent risk factors for major adverse cardiovascular events (MACE) in CHD patients were screened through Lasso regression analysis and Cox regression analysis, and a nomogram prediction model was constructed. The predictive performance of the model was evaluated using time-dependent receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis. Results Variables were screened through Lasso regression and Cox regression analysis. The final model included nine independent predictors, namely age, smoking history, clinical phenotype of CHD, the number of coronary artery lesions, Gensini score, BNP, Lp-PLA2, LP(a), and the history of statin use. The area under the ROC curve in the training set was 0.897, 0.885, and 0.909 at 1, 2, and 3 years, respectively; The area under the ROC curve in the validation set was 0.885, 0.881, and 0.923 at 1, 2, and 3 years, respectively. These results demonstrated that the model had excellent discriminatory power.The calibration curves and decision curves demonstrated that the model had high clinical practicality in predicting the occurrence of MACE in CHD patients. Conclusion The nomogram prediction model based on LP-PLA2, LP(a) and other risk factors provides an effective tool for the prognosis assessment of CHD patients, facilitating the early identification of high-risk patients and enabling individualized intervention.

More+