1.Analysis of HPV Infection Characteristics and Influencing Factors for Lesion Grade in Patients with Cervical Squamous Intraepithelial Lesion and Cervical Cancer
Jingjing HAN ; Lijie ZHANG ; Ruyu CAI ; Haili LI ; He WANG ; Le DANG ; Hongda CHEN ; Ming'e LI ; Lan ZHU
Medical Journal of Peking Union Medical College Hospital 2026;17(1):156-165
To summarize the distribution characteristics of human papillomavirus(HPV) infection types in patients with cervical squamous intraepithelial lesion(SIL) and cervical cancer(CC), and to explore the impact of HPV vaccination, HPV infection types, and general clinical data on different grades of cervical lesions. Clinical data of women attending the gynecological colposcopy clinic of Shenzhen People's Hospital from January 2020 to December 2023 were retrospectively collected. Patients with HPV genotyping records and histopathologically diagnosed SIL or CC were included and divided into three groups based on pathological results: low-grade squamous intraepithelial lesion(LSIL) group, high-grade squamous intraepithelial lesion(HSIL) group, and CC group. The distribution of high-risk HPV subtypes was analyzed among the three groups, and multivariate Logistic regression was used to identify influencing factors for high-grade cervical lesions. A total of 4162 patients were included, comprising 4057 cervical SIL patients(3317 LSIL and 740 HSIL) and 105 CC patients. The overall mean age was(39.9±11.2) years. The HPV infection rate was 95.1%(3959/4162), and 25.0%(1040/4162) of patients had received HPV vaccination. Among high-risk HPV infections, HPV 52, HPV 16, HPV 58, and HPV 18 were the most common subtypes. HPV 52 had the highest infection rate in the LSIL group(27.6%), while HPV 16 was the most prevalent in the HSIL group(45.3%) and CC group(64.9%). Multivariate Logistic regression analysis showed that HPV vaccination( HPV infection is common in patients with SIL and CC, but the distribution of high-risk HPV subtypes varies among different grades of cervical lesions. It is recommended to strengthen cervical cancer screening and monitoring of key high-risk HPV infections in older and multiparous women in Shenzhen, and to continue promoting HPV vaccination.
2.Treatment Principles and Paradigm of Diabetic Microvascular Complications Responding Specifically to Traditional Chinese Medicine
Anzhu WANG ; Xing HANG ; Lili ZHANG ; Xiaorong ZHU ; Dantao PENG ; Ying FAN ; Min ZHANG ; Wenliang LYU ; Guoliang ZHANG ; Xiai WU ; Jia MI ; Jiaxing TIAN ; Wei ZHANG ; Han WANG ; Yuan XU ; .LI PINGPING ; Zhenyu WANG ; Ying ZHANG ; Dongmei SUN ; Yi HE ; Mei MO ; Xiaoxiao ZHANG ; Linhua ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):272-279
To explore the advantages of traditional Chinese medicine (TCM) and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications (DMC), refine key pathophysiological insights and treatment principles, and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine, hosted by the China Association of Chinese Medicine, was held in Beijing, 2024. Experts in TCM, Western medicine, and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis, diagnostic and treatment challenges, and mechanism research related to DMC, ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development, research prioritization, and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM, strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.
3.Association of physical activity and balance ability with disability: a cross-sectional study based on NHANES
Xuejiao HAN ; Jipeng SHI ; Jinpeng WEI ; Tianrui ZHU ; Hongqi XU ; Muhan HE
Chinese Journal of Rehabilitation Theory and Practice 2026;32(1):1-12
ObjectiveTo investigate the association of physical activity and balance ability with disability, and to further examine the mediating role of balance ability in the relationship between physical activity and disability. MethodsBased on data from 2021 to 2023 National Health and Nutrition Examination Survey (NHANES) cycle, a multivariable Logistic regression model was constructed to evaluate the independent association of physical activity and balance ability with disability, with covariates adjusted in four steps. An interaction model was further developed to assess the multiplicative interaction between physical activity and balance ability. Mediation analysis was performed using the Baron and Kenny three-step approach to examine the mediating role of balance ability, and the proportion of the mediation effect was calculated. The significance of the mediation effect was assessed using the Delta method, and robustness was verified through the Bootstrap method with a fixed random seed. Multiple sensitivity analyses were conducted to ensure the stability of the results. ResultsA total of 3 902 participants were included, with a prevalence of disability of 17.7%. Multivariable regression analysis showed that, after adjusting for all covariates, both light-intensity physical activity (LIPA) (OR = 0.489, 95%CI 0.380 to 0.629, P < 0.001) and high-intensity physical activity (HIPA) (OR = 0.493, 95%CI 0.371 to 0.656, P < 0.001) were significantly associated with a reduced risk of disability, whereas impaired balance ability was significantly associated with an increased risk (OR = 1.579, 95%CI 1.266 to 1.970, P < 0.001). The interaction effect analysis showed that the interaction between physical activity and balance ability were not significant (P > 0.05), however, the main effect of LIPA remained robust (β = -0.597, SE = 0.221, OR = 0.550, P = 0.007), while impaired balance ability was significantly associated with an increased risk of disability (β = 0.577, SE = 0.231, OR = 1.780, P = 0.012). The mediation analysis further indicated that balance ability played a robust mediating role in the association between LIPA and disability, with a mediation proportion of 21.1%. The indirect effect (a × b) was statistically significant (P < 0.001), and the 95% confidence intervals derived from the bootstrap method did not include zero. ConclusionPhysical activity and balance ability are significantly associated with disability, and balance ability may mediate the relationship between physical activity and disability, highlighting its potential value in disability risk assessment and intervention strategies.
4.Concentrations characteristics of common air pollutants and health risk assessment of practitioners in hair and beauty salons in Shanghai, 2016–2024
Jiao CHEN ; Tian CHEN ; Xiaoyu WANG ; Yewen SHI ; Fengchan HAN ; Yi HE ; Xiaodong SUN ; Xianliang WANG
Journal of Environmental and Occupational Medicine 2026;43(3):326-332
Background As common public facilities essential to daily life, hair and beauty salons frequently contain various airborne toxic and hazardous pollutants potentially leading to adverse health effects for salon practitioners. Objective To characterize the indoor air pollution profiles of common contaminants in hair and beauty salons in Shanghai and to evaluate the associated health risks for practitioners, in order to provide a scientific basis for strengthening the public health management in Shanghai and protecting the health of practitioners. Methods The air quality monitoring data of hair and beauty salons in Shanghai from 2016 to 2024 were obtained from the “Health Hazard Factors Monitoring Program for Public Places” of the National Institute of Environmental Health, Chinese Center for Disease Control and Prevention. Monitoring indicators included particulate matter ≤10 μm in aerodynamic diameter (PM10), particulate matter ≤2.5 μm in aerodynamic diameter (PM2.5), formaldehyde, ammonia, benzene, toluene, and xylene. Indicator compliance rates were calculated across various years in accordance with GB 9666-1996 Hygienic standard for barber shop and beauty shop and GB 37488-2019 Hygiene indicators and limit for public places; specifically, PM2.5 was assessed against the limits stipulated in GB/T 18883-2022 Standards for indoor air quality. A questionnaire survey was conducted among salon practitioners to collect weekly working days and daily working hours. The non-carcinogenic risks associated with inhalation exposure to formaldehyde, ammonia, benzene, toluene, and xylene as well as the carcinogenic risks posed by formaldehyde and benzene were evaluated following WS/T 777-2021 Technical guide for environmental health risk assessment of chemical exposure and the U.S. Environmental Protection Agency inhalation risk model. Results The overall compliance rates of PM10, formaldehyde, ammonia, benzene, and toluene in the air of hair and beauty salons in Shanghai from 2016 to 2024 were 92.13%, 96.59%, 96.15%, 94.93%, and 94.97%, respectively; the overall compliance rate of xylene was a little lower (85.92%), and the overall compliance rate of PM2.5 was 57.18%. The P50 concentrations of PM10, PM2.5, formaldehyde, ammonia, benzene, toluene, and xylene did not exceed the corresponding limits. The P50 of non-carcinogenic risk indicator (hazard quotient, HQ) for formaldehyde, ammonia, benzene, toluene, and xylene were <1. The probabilities of non-carcinogenic risk HQ >1 for formaldehyde and xylene were 41.4% and 10.9%, respectively, which were higher than that of other pollutants. The P50 of carcinogenic risk (CR) for formaldehyde and benzene were between 1.0×10−6 and 1.0×10−4, while the probabilities of CR >1.0×10−4 were 16.9% and 14.0%, respectively. Conclusion The overall compliance rate of common pollutant concentrations in the air of hair and beauty salons in Shanghai is high, and the hygienic condition meets the requirements of national standards. The non-carcinogenic health risks posed by formaldehyde and xylene to employees (with formaldehyde being more prominent), as well as the carcinogenic risks associated with formaldehyde and benzene, deserve heightened attention in future health supervision.
5.The Role and Regulatory Mechanisms of FOXO1 in Hepatic Lipid Deposition
Meng JIA ; Fang-Hui LI ; Shi-Zhan YAN ; Ai-Ju LI ; Yi-Le WANG ; Pin-Shi NI ; Jia-Han HE ; Yin-Lu LI
Progress in Biochemistry and Biophysics 2026;53(4):905-919
Metabolic associated fatty liver disease (MAFLD) is fundamentally driven by an imbalance in hepatic fatty-acid flux: the influx of fatty acids exceeds the liver’s capacity for disposal, resulting in excessive hepatic lipid accumulation, predominantly in the form of triglycerides (TGs). The occurrence and progression of MAFLD depend on disordered regulation across multiple metabolic steps, including fatty-acid uptake, de novo lipogenesis (DNL), fatty-acid oxidation (FAO), and very low-density lipoprotein (VLDL) export. Forkhead box protein O1 (FOXO1) is a key transcriptional regulator within the hepatic network coordinating glucose and lipid metabolism. Under metabolic stress and insulin resistance (IR), FOXO1 expression is frequently increased, whereas its inhibitory phosphorylation is reduced. These changes enhance FOXO1 nuclear localization and transcriptional activity, thereby reprogramming the expression of genes related to metabolism in the liver. Because hepatic lipid deposition is the central pathological feature of MAFLD, the functional status of FOXO1 directly influences hepatic lipid homeostasis. Growing evidence suggests that FOXO1 can exert bidirectional, environment-dependent effects on hepatic lipid accumulation; however, the molecular basis for this functional switch remains incompletely understood. This review systematically summarizes the biological functions and regulatory mechanisms of FOXO1 and its roles in hepatic lipid metabolism, with a particular focus on its crosstalk with insulin signaling. FOXO1 expression is shaped by RNA modifications and epigenetic regulation mediated by non-coding RNAs. Its transcriptional output is precisely governed by post-translational modifications—such as phosphorylation and acetylation—as well as by coordinated nucleocytoplasmic shuttling. Notably, these regulatory patterns vary markedly across nutritional states, degrees of insulin resistance, and stages of disease. In the fed state, insulin/IGF-1 signaling activates the PI3K-AKT pathway, promoting the inhibitory phosphorylation of FOXO1 and facilitating additional modifications, including acetylation, methylation, and ubiquitination. Together, these events drive FOXO1 export from the nucleus and dampen its transcriptional activity, suppressing gluconeogenesis and constraining lipogenic programs. Conversely, during fasting or when insulin signaling is weakened, FOXO1 inhibition is relieved. FOXO1 accumulates in the nucleus, binds to DNA, and regulates the transcription of downstream target genes. Mechanistically, FOXO1 can aggravate hepatic lipid accumulation by activating genes involved in TG synthesis while repressing FAO-related pathways, thereby favoring storage over oxidation. However, under specific conditions, FOXO1 may also alleviate the hepatic lipid burden by promoting TG hydrolysis and enhancing VLDL secretion, thereby reducing the net hepatic lipid load. In addition, lipotoxic signals mediated by ceramides and diacylglycerols (Cer/DAG) activate atypical protein kinase C (aPKC), further exacerbating the disruption of the AKT-FOXO1 axis. This vicious cycle ultimately produces a metabolic paradox in which increased hepatic glucose output coexists with persistent, insulin-independent lipogenesis, accelerating MAFLD progression. Importantly, FOXO1 regulation is not uniform: during early metabolic overload, insulin-mediated suppression may remain effective, whereas in advanced insulin resistance, the loss of AKT control permits sustained FOXO1 activity. Such stage-dependent dynamics may help explain why FOXO1 can either promote steatosis or, in certain contexts, support programs that facilitate lipid turnover. Accordingly, interventions should be liver-specific and tuned to the disease stage, aiming to curb maladaptive FOXO1 signaling while preserving its capacity to promote triglyceride hydrolysis and VLDL secretion when advantageous. Overall, this review offers an important perspective on MAFLD pathogenesis, emphasizing FOXO1 as a potential therapeutic target and providing a theoretical basis for developing liver-specific, disease-course-dependent precision interventions.
6.Therapeutic efficacy of ruxolitinib combined with low-dose hormone in aGVHD after allogeneic hematopoietic stem cell transplantation
Yue HU ; Xupai ZHANG ; Sihan LAI ; Shan ZHANG ; Lei MA ; Xiao WANG ; Yan DENG ; Ying HAN ; Ying HE ; Guangcui HE ; Hai YI
Chinese Journal of Blood Transfusion 2026;39(4):506-512
Objective: To evaluate the efficacy and safety of ruxolitinib combined with low-dose hormone for patients with acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods: Thirty patients with aGVHD after allo-HSCT admitted to the Department of Hematology of the General Hospital of Western Theater Command from November 2021 to November 2024 were retrospectively analyzed. All patients were treated with low-dose hormone (methylprednisolone 0.3-1 mg kg
-d
) combined with ruxolitinib 5-10 mg d
. The efficacy and adverse reactions were observed during the follow-up period to analyze the survival outcomes of the patients. Results: A total of 30 patients with aGVHD after allo-HSCT were included in this study, consisting of 15 (50%) males and 15 (50%) females with a median age of 34 year-old (ranging from 14 to 62). Classification by disease type: there were 18 cases of acute myeloid leukemia, 4 cases of acute lymphoblastic leukemia, 4 cases of aplastic anemia, and 4 cases of myelodysplastic syndrome. Classification by aGVHD severity: there were 27 cases (90%) of Ⅱ-Ⅳ degree aGVHD and 11 cases (36.7%) of Ⅲ-Ⅳ degree aGVHD. Ruxolitinib in combination with low-dose glucocorticoid treatment yield responses in 28 (93.3%) patients, of which 27 (90%) achieved complete remission (CR), while 1 (3.3%) showed partial remission (PR). One patient (3.3%) had no response (NR), and 1 patient (3.3%) exhibited progressed disease (PD). Overall survival (OS) at 1 year of transplantation was 73.9% (95%CI 49.5% to 87.7%), progression-free survival (PFS) was 93.3% (95%CI 75.9% to 98.3%), non-relapse mortality (NRM) was 20.6% (95%CI 7.9% to 47.4%), and median survival time was 27.6 months. Conclusion: Ruxolitinib combined with low-dose hormones is safe and effective in the treatment of aGVHD after allo-HSCT.
7.Mechanism of Bushen huoxue formula in improving pregnancy outcomes in mice with antiphospholipid antibody-related recurrent spontaneous abortion
Yongmei HAN ; Tianwei CUI ; Jundan XIE ; Xinyu HE ; Yuting GONG ; Manman WANG
China Pharmacy 2026;37(11):1408-1415
OBJECTIVE To investigate the mechanism of Bushen huoxue formula in improving pregnancy outcomes in mice with antiphospholipid antibody-related recurrent spontaneous abortion (aPL-RSA). METHODS SPF female BALB/c mice were randomly divided into normal control group, adjuvant control group, model group, and Bushen huoxue formula group. The aPL-RSA mouse model was established by immunization with β 2 glycoproteinⅠcombined with Freund’s adjuvant. From gestational day 0, the Bushen huoxue formula group was administered 1.653 6 g/(kg·d) of the prescription by gavage, while the other three groups received an equal volume of normal saline, twice daily for 15 consecutive days. The uterine appearance of pregnant mice was observed; individual embryo weight was recorded, and the embryo resorption rate was calculated. The proportion of activated CD41 + CD62p + platelets in platelet-rich plasma (PRP) was detected. Serum levels of thromboxane B2 (TXB2) and platelet factor 4 (PF4) were measured. Morphological changes of placental tissue were observed. The cell apoptosis rate of placental tissue was detected. The levels of malondialdehyde (MDA) content, tot al superoxide dismutase (T-SOD) activity, the interleukin-1β (IL-1β) and IL-18, the protein expressions of matrix metalloproteinase-3 (MMP-3), MMP-9, nuclear proliferation antigen-67 (Ki67), superoxide dismutase 2 (SOD2), Nod-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), and caspase-1, as well as the mRNA expressions of superoxide dismutase 2(SOD2), NLRP3, ASC and caspase-1 of placental tissue were detected. RESULTS Compared with the normal control group and adjuvant control group, the model group showed significant decreases in individual embryo weight, T-SOD activity of placental tissue, and relative expression levels of SOD2 mRNA and protein ( P <0.05); while significant increases were observed in embryo resorption rate, proportion of activated CD41 + CD62p + platelets in PRP, serum levels of TXB2 and PF4, placental cell apoptosis rate, MDA content in placental tissue, relative protein expressions of MMP-3, MMP-9 and Ki67, relative protein and mRNA expression levels of NLRP3, ASC and caspase-1 as well as IL-1β and IL-18 levels ( P <0.05). The model group also exhibited irregular uterine morphology with localized atrophy in some uterine horns; placental tissue showed fragmentation of decidual cells, extensive vacuolization and necrosis of trophoblast cells in the labyrinthine zone, accompanied by vascular reduction. After intervention with Bushen huoxue formula, the above indicators were significantly reversed ( P <0.05), and uterine morphology and pathological damage were markedly improved. CONCLUSIONS Bushen huoxue formula can effectively improve pregnancy outcomes in aPL-RSA model mice, and its mechanism may be closely related to inhibiting platelet activation, reducing oxidative stress injury, enhancing trophoblast cell function, and suppressing the inflammatory response mediated by the NLRP3 inflammasome pathway.
8.Modified Huangqi Jianzhong Decoction Alleviates Gastric Precancerous Conditions in Mice by Regulating Mitochondrial Function via FoxO3/ROS Signaling Pathway
Yueqiang WEN ; Li ZHOU ; Dan LUO ; Maoyuan ZHAO ; Jun HAN ; Xueyi LI ; Jianguo LI ; Zhelin HE ; Tao SHEN ; Jinhao ZENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):216-225
ObjectiveTo investigate the therapeutic effects and mechanisms of modified Huangqi Jianzhong decoction (HQJZ) on gastric precancerous conditions (GPC). MethodsIn the cell experiment, human gastric mucosal epithelial cells underwent malignant transformation induced by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) for the modeling of GPC (MC cells). The cells were allocated into four groups: control , model, low-dose HQJZ (HQJZ-L), and high-dose HQJZ (HQJZ-H). The control and model groups were cultured with the complete medium, while HQJZ-L and HQJZ-H groups received additional interventions with HQJZ at low (0.5 g·L-1) and high (1.0 g·L-1) doses, respectively. Cell counting kit-8 (CCK-8) assay was used to evaluate cytotoxicity, Transwell assay to assess cell invasion, Annexin V-FITC/PI staining to detect apoptosis, immunofluorescence assay to analyze reactive oxygen species (ROS) expression and mitochondrial autophagy, and Western blot to verify expression of proteins in key pathways. In the animal experiment, the GPC model was established in healthy BALB/c mice through MNNG induction. Twenty-four mice were allocated into four groups: control, model, HQJZ-L, and HQJZ-H. Control and model groups received normal saline (10 mL·kg-1·d-1) orally, while HQJZ-L and HQJZ-H groups were administrated with low-dose (6.24 g·kg-1·d-1) and high-dose (12.48 g·kg-1·d-1) HQJZ, respectively. After treatment, hematoxylin‑eosin (HE) staining and AB-PAS staining were performed to observe histopathological changes in the gastric tissue. Immunofluorescence assay was used to detect reactive oxygen species (ROS) and microtubule-associated protein 1 light chain 3 (LC3) levels in the gastric mucosa, TdT-mediated dUTP nick-end labeling (TUNEL) staining to assess apoptosis rates, and Western blotting and immunohistochemistry (IHC) to analyze the expression levels of Ki67, proliferating cell nuclear antigen (PCNA), and foxhead box O3 (FoxO3). ResultsCell viability assays showed that HQJZ dose-dependently reduced MC cell viability compared with the control group (P<0.05, P<0.01). Transwell assays revealed that the model group exhibited enhanced cell invasion compared with the control group (P<0.05). Compared with the model group, HQJZ treatment attenuated the cell invasion (P<0.05). Gastric mucosal pathology in mice demonstrated that compared with the control group, the model group showed elevated HE and AB-PAS pathological scores (P<0.05), while HQJZ treatment reduced these scores (P<0.05). Transmission electron microscopy revealed increased mitochondrial number and volume in the model group compared with the control group. HQJZ treatment resulted in abnormal mitochondrial structure and significant alterations in rough endoplasmic reticulum morphology and distribution, presenting as dilated and hollow forms. Mitochondrial and apoptosis assessments indicated that compared with the control group, the model group exhibited enhanced Mito Tracker Green fluorescence (P<0.05), no significant change in DCFH-DA fluorescence, Mito Tracker Red CMXRos fluorescence, ROS immunofluorescence, or malondialdehyde (MDA) level, increased GSH level (P<0.05), enhanced LC3 fluorescence (P<0.05), no significant change in apoptosis rate, and elevated ATP content in cells and mouse serum (P<0.05). Compared with the model group, HQJZ treatment reduced Mito Tracker Green fluorescence (P<0.05), increased DCFH-DA fluorescence, Mito Tracker Red fluorescence, MDA level, LC3 fluorescence, and apoptosis rate (P<0.05), and decreased cellular ATP content (P<0.05). The HQJZ-L group showed no significant change in ROS immunofluorescence or GSH level, whereas the HQJZ-H group demonstrated enhanced ROS immunofluorescence and glutathione (GSH) level (P<0.05). Immunohistochemistry and Western blotting revealed that compared with the control group, the model group exhibited increased numbers of PCNA- and Ki67-positive cells (P<0.05) and elevated protein levels of FoxO3, sirtuin 1 (SIRT1), and B-cell lymphoma 6 (Bcl-6) (P<0.05). HQJZ treatment reduced the numbers of PCNA- and Ki67-positive cells (P<0.05) and lowered the protein levels of FoxO3, SIRT1, and Bcl-6 (P<0.05). ConclusionHQJZ alleviates the progression of gastric precancerous lesions by regulating mitochondrial function via the FoxO3/ROS pathway and promoting apoptosis of GPC-malignant cells.
9.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.
10.Effect of graphene oxide/hydroxyapatite composite coating on immune activity of RAW264.7 macrophages
Lisha MA ; Huiyu HE ; Wufanbieke·Baheti ; Shangyi LYU ; Xiangzhen HAN
Chinese Journal of Tissue Engineering Research 2025;29(10):2023-2029
BACKGROUND:There are some problems such as bone fusion failure or peri-implant infection after implantation of pure titanium implants.Therefore,surface improvement of titanium implants has become a hot topic in research.Macrophages are an immune defense line of the body in response to external stimuli,and the relevant immune response of any biomaterials implanted in the body is related to macrophages. OBJECTIVE:The graphene oxide/hydroxyapatite composite coating on titanium alloy surface was prepared by electrochemical deposition method.The surface characteristics of the coating and the growth and polarization of macrophage RAW264.7 on the surface were analyzed. METHODS:The graphene oxide coating and graphene oxide/hydroxyapatite composite coating on titanium alloy surface was prepared by electrochemical deposition method.The physical properties of the coating were characterized.Pure titanium sheets(blank group),titanium sheets deposited with pure GO coating(control group)and titanium sheets deposited with graphene oxide/hydroxyapatite composite coating(experimental group)were co-cultured with macrophages RAW264.7,respectively.Cell proliferation was detected by CCK-8 assay and DAPI staining.Cell adhesion on the surface of titanium was observed by scanning electron microscopy,and cell polarization phenotype was detected by flow cytometry. RESULTS AND CONCLUSION:(1)Under scanning electron microscope,the pure titanium sheet showed directional scratches and a few punctate pits.After the pure graphene oxide coating was deposited,the surface of titanium sheet showed more cracks,gullies and particles of uneven size,and the wrinkle-like structure characterized by graphene oxide.After the composite coating was deposited,the surface of the titanium sheet was smooth,and the pellet size was more uniform.The water contact angle of composite coated titanium sheet was lower than that of pure titanium sheet and pure graphene oxide coated titanium sheet(P<0.05).(2)CCK-8 assay and DAPI staining showed that compared with the blank group and the control group,the cell proliferation in the experimental group was faster.Scanning electron microscopy showed that RAW264.7 cells all adhered to the surface of the three groups of titanium sheets.With the extension of culture time,cell morphology changed from round to spindle shape.After 7 days of culture,cells in the blank group extended short and few pseudopods;cells in the control group extended long and more pseudopods,and cells in the experimental group extended short and more pseudopods,and the overall cell fullness in the experimental group was the best.Flow cytometry results showed that the cells in the experimental group showed a higher proportion of M2 polarization in the anti-inflammatory direction.(3)These findings conclude that graphene oxide/hydroxyapatite composite coating has good physical,chemical,and biological properties.

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