1.miRNA-378a overexpression of macrophage cell line composite collagen sponge:anti-inflammation and tissue repair promotion
Sifan WANG ; Huiyu HE ; Quan YANG ; Xiangzhen HAN
Chinese Journal of Tissue Engineering Research 2025;29(4):789-799
BACKGROUND:The regulation of M1/M2 polarization direction of macrophages is particularly critical in tissue engineering applications,and timely regulation can minimize proinflammatory,anti-inflammatory,or tissue healing responses. OBJECTIVE:To implant lentivirus-mediated miRNA-378a macrophage strain complex collagen to detect the expression level of immune regulation in the in vivo environment,and further clarify the influence of miRNA-378a in promoting macrophage M2 polarization in immune regulation and tissue repair in the in vivo environment. METHODS:Lentivirus-mediated miRNA-378a overexpressing macrophage cell lines and negative control virus macrophage lines were amplified and screened,and the macrophage lines were recovered and cultured together with collagen sponge to form a composite scaffold,which was divided into the following groups:(1)Positive group:miRNA-378a overexpressing macrophage-collagen sponge composite;(2)negative group:negative control of virus-mediated miRNA-378a macrophage-collagen sponge composite;(3)control group:macrophage-collagen sponge;(4)blank control group:collagen sponge.The cell density,phenotype,and adhesion of each group were observed by immunofluorescence and scanning electron microscopy.The cells were implanted into the subcutaneous model of the back of mice,and the mice were sacrificed 4 and 7 days after modeling.The direction of macrophage polarization in the collagen sponge composite of macrophages with miRNA-378a overexpression mediated by lentivirus and its effect on immune regulation and tissue repair were analyzed by gross observation,hematoxylin-eosin staining,MASSON staining,and immunohistochemistry. RESULTS AND CONCLUSION:(1)Under immunofluorescence microscopy,the macrophage cell lines in each group were observed to form a composite scaffold with the collagen sponge.(2)Under scanning electron microscope,lentivirus-mediated miRNA-378a macrophages in the positive group proliferated in cell density,had spherical,elliptic and polygonal differentiation,and had more pseudofeet than other groups.(3)Under general observation,the overall 7-day healing was better than that at 4 days.Lentivirus-mediated miRNA-378a macrophages in the positive group healed better than other groups regardless of 4 and 7 days.(4)Lentivirus-mediated miRNA-378a macrophages in the positive group under hematoxylin-eosin staining and MASSON staining had more amounts of fibrocytes,capillaries,fibroblasts,and collagen fiber hyperplasia.(5)Immunohistochemistry showed that lentivirus-mediated miRNA-378a macrophages in the positive group were more positive in 4-and 7-day M2 polarized cells than in other groups.The macrophages of the control and negative groups in 4-and 7-day M2 polarized cells were greater than that of the blank control group.There was no statistical difference between the control group and the negative group.The number of stained cells in the positive,negative,and control groups regardless of 4 and 7 days was higher than that in the blank control group,and the positive group>negative group ≈ control group>blank control group.(6)It is concluded that macrophages with miRNA-378a overexpression have a large amount of fibroblasts,capillaries,fibroblasts,and collagen fiber hyperplasia in vivo,which has a positive effect on tissue repair,and can promote the polarization of macrophages towards M2 type and inhibit the polarization of M1 type,thus contributing to reducing the inflammatory response of the body.
2.Epidemiological characteristics of influenza in Huzhou City from 2014 to 2023
HU Xiaoqiang ; LIU Yan ; ZHOU Sifan ; ZHANG Zizhe ; WANG Yuda ; SHEN Jianyong
Journal of Preventive Medicine 2025;37(9):959-962
Objective:
To analyze the epidemiological characteristics of influenza in Huzhou City, Zhejiang Province from 2014 to 2023, so as to provide a reference for the improvement of influenza prevention and control measures.
Methods:
The data of influenza cases in Huzhou City from 2014 to 2023 were collected from the China Disease Prevention and Control Information System. Descriptive epidemiological methods were used to analyze the population and regional distribution characteristics of influenza. Annual percent change (APC) and average annual percent change (AAPC) were used to analyze the trend of influenza incidence in Huzhou City from 2014 to 2023.
Results:
A total of 83 277 influenza cases were reported in Huzhou City from 2014 to 2023, with an average annual reported incidence of 268.68/105. From 2014 to 2023, the reported incidence of influenza in Huzhou City showed an upward trend (AAPC=68.748%, P<0.05), with a slow upward trend from 2014 to 2021 (APC=31.055%, P<0.05) and a sharp upward trend from 2021 to 2023 (APC=308.782%, P<0.05). The average annual reported incidence of influenza was 270.72/105 in males and 266.54/105 in females, and the difference was not statistically significant (P>0.05). The average annual reported incidence of influenza in children aged 5-<15 years was 1 502.77/100 000. The reported incidences of influenza in Deqing county, Changxing county, and Anji county were 551.44/100 000, 370.47/100 000, and 175.31/100 000, respectively. From 2014 to 2023, the trends of reported influenza incidence in males, females, residents aged 5-<15 years, and 15-<25 years were consistent with the whole population. The reported influenza incidence in each district (county) from 2021 to 2023 was consistent with Huzhou City from 2021 to 2023.
Conclusions
The reported incidence of influenza in Huzhou City showed an overall upward trend from 2014 to 2023, especially from 2021 to 2023. There was no significant gender difference. The majority of the cases were aged 5-<15 years, and the high incidence areas were Deqing County.
3.A simple widely applicable hairy root transformation method for gene function studies in medicinal plants.
Xue CAO ; Zhenfen QIN ; Panhui FAN ; Sifan WANG ; Xiangxiao MENG ; Huihua WAN ; Wei YANG ; Shilin CHEN ; Hui YAO ; Weiqiang CHEN ; Wei SUN
Acta Pharmaceutica Sinica B 2025;15(8):4300-4305
Genetic transformation is a fundamental tool in molecular biology research of medicinal plants. Tailoring transgenic technologies to each distinct medicinal plant would necessitate a substantial investment of time and effort. Here, we present a simple hairy root transformation method that does not require sterile conditions, utilizing Agrobacterium rhizogenes strain K599 and the visible RUBY reporter system. Transgenic hairy roots were obtained for six tested medicinal plant species, roots or rhizomes of which have recognized medicinal value, spanning four botanical families and six genera (Platycodon grandiflorus, Atractylodes macrocephala, Scutellaria baicalensis, Codonopsis pilosula, Astragalus membranaceus, and Glycyrrhiza uralensis). Furthermore, two previously identified Glycyrrhiza uralensis UGTs that convert liquiritigenin into liquiritin in heterologous systems were studied in planta using the method. Our results indicate that overexpression of GuUGT1 but not GuUGT10 and Cas9-mediated knockout of GuUGT1 profoundly influenced the accumulation of liquiritin and isoliquiritin in licorice roots. Therefore, the method described here represents a simple, rapid and widely applicable hairy root transformation method that enables fast gene functional study in medicinal plants.
4.TPMGD: A genomic database for the traditional medicines in Pakistan.
Rushuang XIANG ; Huihua WAN ; Wei SUN ; Baozhong DUAN ; Weiqian CHEN ; Xue CAO ; Sifan WANG ; Chi SONG ; Shilin CHEN ; Yan WANG ; Atia-Tul WAHAB ; M IQBAL CHOUDHARY ; Xiangxiao MENG
Chinese Herbal Medicines 2025;17(1):87-93
OBJECTIVE:
In Pakistan, traditional medicines are an important component of the medical system, with numerous varieties and great demands. However, due to the scattered resources and the lack of systematic collection and collation, adulteration of traditional Pakistani medicine (TPM) is common, which severely affects the safety of their medicinal use and the import and export trades. Therefore, it is urgent to systematically organize and unify the management of TPM and establish a set of standards and operable methods for the identification of TPM.
METHODS:
We collected and organized the information on 128 TPMs with regard to their medicinal parts, efficacy, usage, and genetic material, based on Pakistan Hamdard Pharmacopoeia of Eastern Medicine: Pharmaceutical Codex. The genetic information of TPM is summarized from national center for biotechnology information (NCBI) and global pharmacopoeia genome database (GPGD). Furthermore, we utilized bioinformatics technology to supplement the chloroplast genome (cp-genome) data of 12 TPMs. To build the web server, we used the Linux + Apache + MySQL + PHP (LAMP) system and constructed the webpage on a PHP: Hypertext Preprocessor (PHP) model view controller (MVC) framework.
RESULTS:
We constructed a new genomic database, the traditional Pakistani medicine genomic database (TPMGD). This database comprises five entries, namely homepage, medicinal species, species identification, basic local alignment search tool (BLAST), and download. Currently, TPMGD contains basic profiles of 128 TPMs and genetic information of 102 TPMs, including 140 cytochrome c oxidase subunit I (COI) sequences and 119 mitochondrial genome sequences from Bombyx mori, 1 396 internal transcribed spacer 2 (ITS2) sequences and 1 074 intergenic region (psbA-trnH) sequences specific to 92 and 83 plant species, respectively. Additionally, TPMGD includes 199 cp-genome sequences of 82 TPMs.
CONCLUSION
TPMGD is a multifunctional database that integrates species description, functional information inquiry, genetic information storage, molecular identification of TPM, etc. The database not only provides convenience for TPM information queries but also establishes the scientific basis for the medication safety, species identification, and resource protection of TPM.
5.Metabolomics as an emerging tool for the pharmacological and toxicological studies on Aconitum alkaloids.
Han DING ; Yamin LIU ; Sifan WANG ; Yuqi MEI ; Linnan LI ; Aizhen XIONG ; Zhengtao WANG ; Li YANG
Chinese Journal of Natural Medicines (English Ed.) 2025;23(2):182-190
Aconitum (Ranunculaceae) has a long-standing history in traditional Chinese medicine (TCM), where it has been widely used to treat conditions such as rheumatoid arthritis (RA), myocardial infarction, and heart failure. However, the potency of Aconitum alkaloids, the primary active components of Aconitum, also confers substantial toxicity. Therefore, assessing the efficacy and toxicity of these Aconitum alkaloids is crucial for ensuring clinical effectiveness and safety. Metabolomics, a quantitative method for analyzing low-molecular-weight metabolites involved in metabolic pathways, provides a comprehensive view of the metabolic state across multiple systems in vivo. This approach has become a vital investigative tool for facilitating the evaluation of their efficacy and toxicity, identifying potential sensitive biomarkers, and offering a promising avenue for elucidating the pharmacological and toxicological mechanisms underlying TCM. This review focuses on the applications of metabolomics in pharmacological and toxicological studies of Aconitum alkaloids in recent years and highlights the significant role of metabolomics in exploring compatibility detoxification and the mechanisms of TCM processing, aiming to identify more viable methods for characterizing toxic medicinal plants.
Aconitum/metabolism*
;
Metabolomics/methods*
;
Alkaloids/metabolism*
;
Humans
;
Animals
;
Drugs, Chinese Herbal/pharmacology*
;
Medicine, Chinese Traditional
6.A high-throughput plant canopy leaf area index inversion model based on UAV-LiDAR.
Yuming LIANG ; Xueyan FAN ; Muqing ZHANG ; Wei YAO ; Xiuhua LI ; Zeping WANG ; Sifan DONG ; Xuechen LI
Chinese Journal of Biotechnology 2025;41(10):3817-3827
To explore the feasibility of using UAV-LiDAR for measuring the leaf area index (LAI) of crop canopies, we employed UAV-LiDAR to scan sugarcane canopies during the tillering and elongation stages, acquiring canopy point cloud data. Subsequently, features such as average row height, projected row area, point cloud density at different canopy layers, and the ratios between these parameters were extracted. Three feature selection methods-partial least squares regression (PLSR), XGBoost feature importance (XGBoost-FI), and random forest-recursive feature elimination (RF-RFE)-were adopted to evaluate and identify the optimal input variables for modeling. With these selected variables, LAI inversion models were developed based on random forest (RF) and adaptive boosting (AdaBoost) algorithms, and their performance was assessed. Among the extracted features, the projected row area Sp and the total row point count Ctotal exhibited strong correlations with LAI, with correlation coefficients of 0.73 and 0.72, respectively. The AdaBoost-based LAI inversion model, using the projected row area Sp, average height Havg, mid-layer point cloud density Cm, and total row point count Ctotal as input variables, achieved the best performance, with a coefficient of determination (Rv²) of 0.713 and a root mean square error (RMSEv) of 0.25 on the validation set. This study provides an effective method for high-throughput acquisition of LAI in field crops, offering valuable scientific support for sugarcane field management and breeding efforts.
Plant Leaves/growth & development*
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Saccharum/growth & development*
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Algorithms
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Unmanned Aerial Devices
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Remote Sensing Technology/methods*
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Crops, Agricultural/growth & development*
7.Risk factor research and risk prediction model establishment for early Q-T interval prolongation after acute myocardial infarction
Sifan LI ; Ying XIAO ; Dongbo WANG ; Sining LIU ; Yadong TANG ; Xuefeng TIAN
Chinese Journal of Primary Medicine and Pharmacy 2024;31(2):175-179
Objective:To establish a prediction model of risk factors for early Q-T interval prolongation after acute myocardial infarction (AMI), which helps prevent and reduce the occurrence of acute malignant events.Methods:This is a case-control study. A total of 100 patients with Q-T interval prolongation after AMI who received treatment at Heilongjiang Provincial Hospital from January 2018 to December 2022 were included in this study. An additional 100 patients without Q-T interval prolongation after AMI who concurrently received treatment in the same hospital were also included in this study. Two model groups, including model group 1 (with Q-T interval prolongation, n = 50) and model group 2 (without Q-T interval prolongation, n = 50), and two test groups, including test group 1 (with Q-T interval prolongation, n = 50) and test group 2 (without Q-T interval prolongation, n = 50), were designated. Logistic regression analysis was performed to construct a prediction model of risk factors for Q-T interval prolongation. The area under the receiver operating characteristic curve was determined to evaluate the prediction model. The value of the prediction model was validated in the test groups. Results:Multivariate logistic regression showed that female gender ( OR = 2.307, 95% CI: 0.09-0.91, P = 0.041) and heart failure ( OR = 3.087, 95% CI: 1.15-8.27, P = 0.025) were independent risk factors for early Q-T interval prolongation after AMI. The area under the receiver operating characteristic curve of the prediction model was 0.770, with a sensitivity of 84.0%, a specificity of 66.0%, the Jordan index of 0.44, and the corresponding optimal critical value of 0.43. This indicates good fit of the model. Conclusion:Female gender and heart failure are independent risk factors for early Q-T interval prolongation after AMI. The model constructed based on the above-mentioned risk factors fits well and has a high predictive value, which helps reduce the occurrence of early Q-T interval prolongation after AMI.
8.Overexpression of miR-378a promotes macrophage M2 polarization and inhibits M1 polarization
Quan YANG ; Huiyu HE ; Sifan WANG ; Shangyi LYU ; Qiqi ZHOU ; Xiangzhen HAN
Chinese Journal of Tissue Engineering Research 2024;28(13):2036-2041
BACKGROUND:M2 macrophages have the function of reducing inflammatory factors and promoting tissue healing.Therefore,how to regulate M2 polarization of macrophages has been a hot research topic in recent years,and some miRNAs have been found to have this function. OBJECTIVE:To investigate the effects of miR-378a on the polarization of the Raw264.7 macrophage cell line. METHODS:The M1 polarization of macrophages was induced by lipopolysaccharide and interferon-γ.Interleukin-4 induced M2 polarization and the expression of endogenous miR-378a in each cell type was detected using qRT-PCR to verify whether miR-378a was involved in the polarization of macrophages.By transfection with lentivirus as the vector of overexpression of miR-378a,the stable expression of miR-378a cell lines was screened.Macrophage M1 polarization was induced synergically by lipopolysaccharide and interferon-γ.Macrophage M2 polarization was induced by interleukin-4.The levels of M1/M2 polarization-related cytokines in the supernatant of the macrophage culture medium were determined by enzyme-linked immunosorbent assay.qRT-PCR was used to detect the polarization characteristics of M1/M2-type macrophages and the mRNA expression levels of related cytokines. RESULTS AND CONCLUSION:(1)The expression level of endogenous miR-378a in Raw264.7 cells of each group increased after macrophage polarization.(2)Compared with the non-transfected group,the expressions of proinflammatory cytokine-induced nitric oxide synthase,tumor necrosis factor-α,interleukin-6 and interleukin-1β in macrophage M1 induced polarization were significantly decreased in the miR-378a transfection group(P<0.05);the levels of inducible nitric oxide synthase,tumor necrosis factor-α and interleukin-6 in cell supernatant were also significantly decreased(P<0.05).(3)Compared with the non-transfected group,the expressions of CD206,interleukin-10 and arginase-I in macrophage M2 induced polarization were significantly increased(P<0.05);the levels of CD206 and interleukin-10 in cell supernatant were also significantly increased(P<0.05)in the miR-378a transfection group.(4)It is indicated that overexpression of miR-378a promotes the M2 polarization of macrophages and inhibits the M1 polarization of macrophages.
9. Mechanisms of Radix Tetrastigma on anti rheumatoid arthritis via regulating the balance of Th17/Treg
Tiefeng WANG ; Wenrui ZHU ; Jie SU ; Rongshuang XIA ; Sifan LI ; Zhaohuan LOU ; Weidong WANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2023;28(2):138-146
AIM: To explore the possible mechanism of Radix Tetrastigma (RT) on anti-rheumatoid arthritis (RA). METHODS: The rat model of RA was established by intradermal injection of complete Freund]s adjuvant into the right hind foot of SD rats. RT Extract with different dosage was continuous intragastric administration for 3 weeks, then, the degree of foot swelling, arthritis index score, joint heat and grip of each rat was measured respectively. ELISA was used to detect the expression levels of Interleukin (IL) 6, IL-17, IL-10, tumor necrosis factor-α, and rheumatoid factor. Fully automatic hemorheometer was applied to measure hemorheology indexes. The number of CD4
10.Changes of complement fragment C3a and its receptor in lung injury induced by silica nanoparticles
Sifan ZHOU ; Zhoujian WANG ; Lei GAO ; Yuansheng DUAN ; Muyue CHEN ; Jiaxiang ZHANG ; Qixing ZHU
Journal of Environmental and Occupational Medicine 2023;40(8):877-883
Background Silica nanoparticles (SiNPs) enter the human body through respiratory tract, digestive tract, and skin, causing body damage. Lung is one of the main damaged organs. Objective To observe the expressions of complement activated fragment C3a and its receptor C3aR in the lungs of mice exposed to SiNPs through respiratory tract, and to explore the involvement of C3a/C3aR in lung injury induced by SiNPs exposure. Methods The ultrastructure of SiNPs (particle size 5-20 nm) was determined under a transmission electron microscope, and the hydrodynamic diameter and surface Zeta potential of SiNPs were determined using a nanoparticle size analyzer. A total of 88 SPF C57BL/6J mice were randomly divided into five groups: a blank control group without any treatment (14 mice), a vehicle control group treated with 50 μL stroke-physiological saline solution by intratracheal instillation (14 mice), and three SiNPs exposure groups (low-dose group, medium-dose group, and high-dose group with 20 mice in each group, who were given 50 μL SiNPs suspension of 7, 21, and 35 mg·kg−1 respectively and exposed once every 3 days for 5 times). The mice were anesthetized on day 1 (1-day model group) and day 15 (15-day model group) after exposure, then sacrificed after extraction of bronchoalveolar lavage fluid (BALF), and lung tissues were retained. The morphological changes of lung tissues were observed by HE staining, the expression level of C3a in BALF was detected by enzyme-linked immunosorbent assay, the deposition of C3a and C3aR in lung tissues were observed by immunohistochemistry, the protein expression level of C3aR was determined by Western blotting, and the localization and semi-quantitative detection of C3a and C3aR in lung tissues was observed by immunofluorescence. Results SiNPs agglomerated in stroke-physiological saline solution. The average hydrodynamic diameter was (185.60±7.39) nm and the absolute value of Zeta potential was (43.33±0.76) mV. The condition of mice in the 1-day model group and the 15-day model group was good, while 2 mice died in the medium-dose group of the 1-day model group due to misoperation. The autopsy results of the two mice showed congestion of the lung tissue, emphysema, and no imperfection of trachea integrity. No death was observed in other dose groups. The HE staining results showed pathological damage to the mouse lung, including alveolar wall thickening and inflammatory cell infiltration after SiNPs exposure. The pathological damage became more serious with the increase of dose. Regarding pathological changes, the 15-day model group was slightly relieved compared with the 1-day model group, but there were still pathological changes. The enzyme-linked immunosorbent assay results showed that there was no difference in the expression level of C3a between the blank control group and the vehicle control group (P>0.05), the expression levels of C3a in the medium-dose group and the high-dose group were significantly higher than that in the vehicle control group (P<0.05). The immunohistochemistry results showed that C3a deposition was consistent with the enzyme-linked immunosorbent assay results. The Western blotting and the immunohistochemistry results showed that C3aR expression was low in the blank control group and the vehicle control group, while the expression in each dose group tended to increase with the increase of dose. The immunofluorescence results showed that the fluorescence signals of C3a and C3aR were weak in the blank control group and the vehicle control group in the 1-day model group and the 15-day model group, while the fluorescence signals in the lung tissues of mice in the SiNPs exposure groups tended to increase with the increase of dose. Conclusion The increased expressions of C3a and C3aR in complement activation may be related to lung injury induced by intratracheal instillation of SiNPs, suggesting that C3a/C3aR may be involved in lung injury induced by SiNPs exposure.


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