Objective To observe the clinical efficacy of Bushen Yiliu Formula(mainly composed of Epimedii Folium,Ligustri Lucidi Fructus,Ecliptae Herba,and Paridis Rhizoma)in treating advanced hormone-sensitive prostate cancer accompanied by bone metastasis of kidney deficiency type.Methods A prospective randomized controlled trial was conducted on 45 patients with advanced hormone-sensitive prostate cancer accompanied by bone metastasis of kidney deficiency type,who received treatment at Guangdong Provincial Hospital of Chinese Medicine from January 2022 to February 2024.The patients were randomly divided into an observation group(23 cases)and a control group(22 cases).All patients received complete androgen blockade therapy combined with bone-protective therapy.Additionally,the observation group was treated with the modified Bushen Yiliu Formula for 6 months.The clinical efficacy of Bushen Yiliu Formula was evaluated with the disease control rate(DCR)and objective response rate(ORR)after 3 and 6 months of treatment,and with the changes in the TCM syndrome scores,Karnofsky Performance Status(KPS)scores,Numerical Rating Scale(NRS)pain scores,and serum levels of total prostate-specific antigen(tPSA)and alkaline phosphatase(ALP)before treatment and after 3 and 6 months of treatment.Changes in serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),and creatinine(Scr)levels before treatment and after 3 and 6 months of treatment were observed to assess the clinical safety of the formula.Results(1)After 3 months of treatment,both groups had no complete remission cases.After 6 months,the observation group hand only one complete remission case.After 3 months of treatment,the ORR and DCR in the observation group were 21.74%(5/23)and 95.65%(22/23),respectively,compared to 22.73%(5/22)and 95.45%(21/22)in the control group.The intergroup comparison showed that there were no significant differences between the two groups(P＞0.05).After 6 months of treatment,the ORR and DCR in the observation group were 60.87%(14/23)and 95.65%(22/23),respectively,significantly higher than those in the control group[22.73%(5/22)and 68.18%(15/22),respectively],with statistically significant differences(P＜0.05).(2)After 3 and 6 months of treatment,TCM syndrome scores and NRS pain scores in both groups were significantly decreased(P＜0.01),and KPS scores were significantly increased(P＜0.01).Except for TCM syndrome scores after 3 months of treatment,the observation group had stronger effect on decreasing TCM syndrome scores and NRS pain scores and on increasing KPS scores after 3 and 6 months of treatment than the control group(P＜0.01).(3)The serum level of tumor-related marker tPSA after 3 months of treatment,and serum tPSA and ALP levels after 6 months of treatment were significantly decreased in both groups compared to the baseline level(P＜0.05 or P＜0.01).The observation group had stronger effect on decreasing serum tPSA level after 3 and 6 months compared to the control group(P＜0.05 or P＜0.01).However,no significant differences of serum ALP level were observed between the two groups after 3 and 6 months of treatment(P＞0.05).(4)Serum levels of safety indicators of ALT,AST,and Scr showed no significant changes in either group after 3 or 6 months of treatment compared to the baseline level(P＞0.05).Conclusion Bushen Yiliu Formula combined with endocrine therapy exerts certain efficacy for the treatment of advanced hormone-sensitive prostate cancer accompanied by bone metastasis of kidney deficiency type.The combined therapy is effective in reducing tumor burden of bone metastases,improving objective response rates,alleviating clinical symptoms,decreasing NRS pain scores,enhancing quality of life,and effectively lowering serum tPSA and ALP levels.Its efficacy is significantly superior to endocrine therapy alone.

Noise-induced hearing loss is a worldwide public health issue that is characterized by temporary or permanent changes in hearing sensitivity. This condition is closely linked to inflammatory responses, and interventions targeting the inflammatory gene tumor necrosis factor-alpha (TNFα) are known to mitigate cochlear noise damage. TNFα-induced proteins (TNFAIPs) are a family of translucent acidic proteins, and TNFAIP6 has a notable association with inflammatory responses. To date, there have been few reports on TNFAIP6 levels in the inner ear. To elucidate the precise mechanism, we generated transgenic mouse models with conditional knockout of Tnfaip6 (Tnfaip6 cKO). Evaluation of hair cell morphology and function revealed no significant differences in hair cell numbers or ribbon synapses between Tnfaip6 cKO and wild-type mice. Moreover, there were no notable variations in hair cell numbers or hearing function in noisy environments. Our results indicate that Tnfaip6 does not have a substantial impact on the auditory system.
Animals ; Mice, Knockout ; Hair Cells, Auditory, Inner/pathology* ; Mice ; Mice, Transgenic ; Hearing Loss, Noise-Induced ; Evoked Potentials, Auditory, Brain Stem/physiology*

Objective·To explore the function and potential mechanism of suppressor of zeste 12(SUZ12)in hepatocellular carcinoma(HCC).Methods·The expression of SUZ12 in HCC patients was analyzed using R language in liver cancer datasets,and relevant survival curves were drawn.Stable knockdown of SUZ12 was established in the liver cancer cell lines LM3 and Huh7.The knockdown efficiency of SUZ12 was assessed using quantitative real-time PCR(qPCR)and Western blotting.Cell proliferation ability was assessed using CCK-8 assay and colony formation assay.Using the hydrodynamic tail vein injection(HTVI)method,Suz12 was knocked out in the livers of fully immunocompetent mice to explore its tumorigenic function in vivo.The molecular mechanism of SUZ12 regulating HCC was explored using The Cancer Genome Atlas(TCGA)database.R language was used to analyze the relationship between SUZ12 and the expression of cancer stem cell(CSC)markers as well as key glycolysis-related genes.Findings were validated in liver cancer cell lines and mouse tumor tissues.Results·The expression of SUZ12 in liver cancer tissues was higher than in adjacent non-tumor tissues,and its expression increased with higher tumor stage.HCC patients with high SUZ12 expression had poorer prognoses.In LM3 and Huh7 liver cancer cell lines,stable knockdown of SUZ12 reduced cell proliferation ability.In the de novo MYC/Trp53-/-mouse liver cancer model,tumor nodule number and size,and tumor burden in liver tissue were reduced after endogenous knockout of Suz12.TCGA analysis showed that high SUZ12 expression in HCC was enriched in multiple tumor proliferation-and metabolism-related pathways.The expression of SUZ12 was positively correlated with CSC markers and key genes in glycolysis pathway.The mRNA levels of CSC markers and key genes in glycolysis pathway were decreased in liver cancer cell lines with stable SUZ12 knockdown and Suz12 knockout mouse HCC tissues.Conclusion·The expression of SUZ12 is significantly increased in HCC patients and is associated with poor prognosis.Stable knockdown of SUZ12 weakens the proliferative ability of liver cancer cells.Knockout of Suz12 in mice in vivo can suppress the occurrence and development of HCC.The high expression of SUZ12 maintains the CSC pool,induces metabolic reprogramming,and promotes the occurrence and progression of HCC.SUZ12 can serve as a potential biomarker for poor prognosis and a novel target for potential therapeutic intervention in HCC.

Objective·To explore the function and potential mechanism of suppressor of zeste 12(SUZ12)in hepatocellular carcinoma(HCC).Methods·The expression of SUZ12 in HCC patients was analyzed using R language in liver cancer datasets,and relevant survival curves were drawn.Stable knockdown of SUZ12 was established in the liver cancer cell lines LM3 and Huh7.The knockdown efficiency of SUZ12 was assessed using quantitative real-time PCR(qPCR)and Western blotting.Cell proliferation ability was assessed using CCK-8 assay and colony formation assay.Using the hydrodynamic tail vein injection(HTVI)method,Suz12 was knocked out in the livers of fully immunocompetent mice to explore its tumorigenic function in vivo.The molecular mechanism of SUZ12 regulating HCC was explored using The Cancer Genome Atlas(TCGA)database.R language was used to analyze the relationship between SUZ12 and the expression of cancer stem cell(CSC)markers as well as key glycolysis-related genes.Findings were validated in liver cancer cell lines and mouse tumor tissues.Results·The expression of SUZ12 in liver cancer tissues was higher than in adjacent non-tumor tissues,and its expression increased with higher tumor stage.HCC patients with high SUZ12 expression had poorer prognoses.In LM3 and Huh7 liver cancer cell lines,stable knockdown of SUZ12 reduced cell proliferation ability.In the de novo MYC/Trp53-/-mouse liver cancer model,tumor nodule number and size,and tumor burden in liver tissue were reduced after endogenous knockout of Suz12.TCGA analysis showed that high SUZ12 expression in HCC was enriched in multiple tumor proliferation-and metabolism-related pathways.The expression of SUZ12 was positively correlated with CSC markers and key genes in glycolysis pathway.The mRNA levels of CSC markers and key genes in glycolysis pathway were decreased in liver cancer cell lines with stable SUZ12 knockdown and Suz12 knockout mouse HCC tissues.Conclusion·The expression of SUZ12 is significantly increased in HCC patients and is associated with poor prognosis.Stable knockdown of SUZ12 weakens the proliferative ability of liver cancer cells.Knockout of Suz12 in mice in vivo can suppress the occurrence and development of HCC.The high expression of SUZ12 maintains the CSC pool,induces metabolic reprogramming,and promotes the occurrence and progression of HCC.SUZ12 can serve as a potential biomarker for poor prognosis and a novel target for potential therapeutic intervention in HCC.

OBJECTIVE To investigate of the regulatory effects of Budi Shenlian Recipe on gut microbiota and the phenotypic transition of tumor-associated macrophages(TAMs)in colorectal cancer(CRC)mice.METHODS Forty male BALB/c mice were divided into blank control group,model group,positive control group,high-dose Budi Shenlian Recipe group,and low-dose Budi Shenlian Recipe group(n=8).CRC-bearing models were established by subcutaneous injection of CT26 cells.Additionally,10 male BALB/c mice were divided into the Budi Shenlian Recipe fecal microbiota transplantation(BFMT)group and model fecal microbiota transplantation(MFMT)group(n=5).The gut microbiota of these mice was cleared using a mixed solution of quadruple antibiotics,followed by subcutaneous injection of CT26 cells to construct pseudo-germ-free CRC-bearing mice.Fecal samples from the model group and high-dose Budi Shenlian Recipe group were collected to prepare fecal microbiota solutions.The BFMT group received gavage with fecal microbiota solution from the high-dose Budi Shenlian Recipe group,while the MFMT group received gavage with fecal micro-biota solution from the model group.Tumor volume changes were observed and recorded.HE staining was used to assess pathological changes in tumor tissues.16S sequencing was performed to analyze changes in gut microbiota.Flow cytometry and immunofluorescence staining were used to evaluate the proportion of M1/M2 type TAMs in tumor tissues.ELISA was used to detect differences in TNF-α and IL-10 levels in tumor tissues.RESULTS Compared to the model group,the tumor volume of mice in the positive control group,high-dose Budi Shenlian Recipe group,and low-dose Budi Shenlian Recipe group grew more slowly(P<0.01).HE staining showed necrotic areas in tumor tissues and reduced mitotic figures in the positive control and Budi Shenlian Recipe groups compared to the model group.16S rRNA sequencing showed no significant differences in Chao1 and ACE indices between the high-dose Budi Shenlian Recipe group and the model group.PCoA analysis indicated a distinct microbial community structure between the blank group and mod-el group,with the microbial structure of CRC mice in the Budi Shenlian Recipe group closer to that of the blank group.Compared to the blank group,the model group showed a significant decrease in the proportion of Muribaculaceae,Muribaculum,Alloprevotella,and Prevotellaceae_UCG-001,and a significant increase in Lachnospiraceae_NK4A136_group,Bacteroides,and Helicobacter.After admin-istering Budi Shenlian Recipe to CRC mice,the community structure of some mice partially reverted to the level of the blank group.Transcriptome sequencing revealed that the most significant biological process(BP)among upregulated genes was the negative regulation of macrophage migration,suggesting that Budi Shenlian Recipe can reduce macrophage migration.Moreover,compared to the model group,the proportion of TAMs cells in the tumor tissues of the Budi Shenlian Recipe group significantly decreased(P<0.001).Simultaneously,compared to the model group,the proportion of M1 type TAMs in the Budi Shenlian Recipe group significant-ly increased(P<0.000 1),while the proportion of M2 type TAMs significantly decreased(P<0.05).Immunofluorescence analysis showed the same trend as flow cytometry.The content of TNF-α in tumor tissues of the Budi Shenlian Recipe group significantly in-creased(P<0.001),and IL-10 content significantly decreased(P<0.001).Additionally,compared to the MFMT group,the tumor volume in the BFMT group grew more slowly(P<0.0001).HE staining showed increased necrotic areas,sparser cell arrangement,and reduced pathological mitosis in the BFMT group.Furthermore,compared to the MFMT group,the proportion of TAMs cells in the tumor tissues of the BFMT group significantly decreased(P<0.01).Compared to the MFMT group,the proportion of M1 type TAMs cells in the BFMT group increased(P<0.000 1),while the proportion of M2 type TAMs cells decreased(P<0.01).Immunofluores-cence analysis further confirmed that Budi Shenlian Recipe fecal microbiota transplantation can reduce the proportion of TAMs in the tumor tissues of CRC mice and promote the conversion from M2 to M1 type,thereby reducing immune suppression in the tumor micro-environment.CONCLUSION Budi Shenlian Recipe can improve gut microbiota dysbiosis in CRC mice and exert anti-CRC effects by reducing tumor infiltration of TAMs and modulating the phenotypic transition of TAMs.There may be a certain correlation between these two effects.

Objective:To explore the association of blood and urinary cadmium levels with lipid profile levels and dyslipidemia in Chinese adults aged 18 to 79 years.Methods:Based on the China National Human Biomonitoring (CNHBM) program, a cross-sectional survey was conducted from 2017 to 2018 using a multi-stage stratified random sampling method, including a total of 10 713 adults aged 18 to 79 years. Data was obtained through questionnaires, physical examinations, biological sample collection, and laboratory testing. Multiple linear mixed effect model (MLMM) and generalized linear mixed effect model (GLMM) were used to analyze the association of blood and creatinine-corrected urinary cadmium levels with lipid profile levels as well as dyslipidemia among adults.Results:The age of 10 713 participants was (47.23±0.24) years, with 5 372 males accounting for 61.3% of the national population. The weighted mean±standard error (SE) of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) was (5.21±0.03), (1.86±0.03), (2.96±0.03), and (1.43±0.01) mmol/L, respectively. The prevalence rate of hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, low HDL-C, and high LDL-C was 16.0%, 21.6%, 6.6%, 13.5%, and 10.0%, respectively. MLMM showed that, after adjusting for relevant confounders, log-transformed blood cadmium levels were positively associated with increased levels of TC, TG and LDL-C ( P<0.05). When blood cadmium levels were categorized into quartiles, compared to the lowest exposure group ( Q1), participants in the highest blood cadmium exposure group ( Q4) had increases of 0.19 (95% CI: 0.06, 0.32) mmol/L in TC and 0.25 (95% CI: 0.08, 0.43) mmol/L in TG. GLMM indicated that, after adjusting for confounders, higher blood cadmium exposure levels were associated with increased risks of hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, and high LDL-C ( P<0.05). Further analysis by quartiles showed that, compared to the blood cadmium Q1 exposure group, the OR value (95% CI) for the Q4 group was 1.53 (1.12, 2.08) for hypercholesterolemia, 1.54 (1.09, 2.17) for hypertriglyceridemia, 2.24 (1.47, 3.40) for mixed hyperlipidemia, and 1.49 (1.07, 2.09) for high LDL-C. Conclusion:The cadmium internal exposure levels are associated with blood lipid profile levels as well as the incidence of dyslipidemia in Chinese adults aged 18 to 79.

Objective:To explore the association of blood and urinary cadmium levels with lipid profile levels and dyslipidemia in Chinese adults aged 18 to 79 years.Methods:Based on the China National Human Biomonitoring (CNHBM) program, a cross-sectional survey was conducted from 2017 to 2018 using a multi-stage stratified random sampling method, including a total of 10 713 adults aged 18 to 79 years. Data was obtained through questionnaires, physical examinations, biological sample collection, and laboratory testing. Multiple linear mixed effect model (MLMM) and generalized linear mixed effect model (GLMM) were used to analyze the association of blood and creatinine-corrected urinary cadmium levels with lipid profile levels as well as dyslipidemia among adults.Results:The age of 10 713 participants was (47.23±0.24) years, with 5 372 males accounting for 61.3% of the national population. The weighted mean±standard error (SE) of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) was (5.21±0.03), (1.86±0.03), (2.96±0.03), and (1.43±0.01) mmol/L, respectively. The prevalence rate of hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, low HDL-C, and high LDL-C was 16.0%, 21.6%, 6.6%, 13.5%, and 10.0%, respectively. MLMM showed that, after adjusting for relevant confounders, log-transformed blood cadmium levels were positively associated with increased levels of TC, TG and LDL-C ( P<0.05). When blood cadmium levels were categorized into quartiles, compared to the lowest exposure group ( Q1), participants in the highest blood cadmium exposure group ( Q4) had increases of 0.19 (95% CI: 0.06, 0.32) mmol/L in TC and 0.25 (95% CI: 0.08, 0.43) mmol/L in TG. GLMM indicated that, after adjusting for confounders, higher blood cadmium exposure levels were associated with increased risks of hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, and high LDL-C ( P<0.05). Further analysis by quartiles showed that, compared to the blood cadmium Q1 exposure group, the OR value (95% CI) for the Q4 group was 1.53 (1.12, 2.08) for hypercholesterolemia, 1.54 (1.09, 2.17) for hypertriglyceridemia, 2.24 (1.47, 3.40) for mixed hyperlipidemia, and 1.49 (1.07, 2.09) for high LDL-C. Conclusion:The cadmium internal exposure levels are associated with blood lipid profile levels as well as the incidence of dyslipidemia in Chinese adults aged 18 to 79.

OBJECTIVE To investigate of the regulatory effects of Budi Shenlian Recipe on gut microbiota and the phenotypic transition of tumor-associated macrophages(TAMs)in colorectal cancer(CRC)mice.METHODS Forty male BALB/c mice were divided into blank control group,model group,positive control group,high-dose Budi Shenlian Recipe group,and low-dose Budi Shenlian Recipe group(n=8).CRC-bearing models were established by subcutaneous injection of CT26 cells.Additionally,10 male BALB/c mice were divided into the Budi Shenlian Recipe fecal microbiota transplantation(BFMT)group and model fecal microbiota transplantation(MFMT)group(n=5).The gut microbiota of these mice was cleared using a mixed solution of quadruple antibiotics,followed by subcutaneous injection of CT26 cells to construct pseudo-germ-free CRC-bearing mice.Fecal samples from the model group and high-dose Budi Shenlian Recipe group were collected to prepare fecal microbiota solutions.The BFMT group received gavage with fecal microbiota solution from the high-dose Budi Shenlian Recipe group,while the MFMT group received gavage with fecal micro-biota solution from the model group.Tumor volume changes were observed and recorded.HE staining was used to assess pathological changes in tumor tissues.16S sequencing was performed to analyze changes in gut microbiota.Flow cytometry and immunofluorescence staining were used to evaluate the proportion of M1/M2 type TAMs in tumor tissues.ELISA was used to detect differences in TNF-α and IL-10 levels in tumor tissues.RESULTS Compared to the model group,the tumor volume of mice in the positive control group,high-dose Budi Shenlian Recipe group,and low-dose Budi Shenlian Recipe group grew more slowly(P<0.01).HE staining showed necrotic areas in tumor tissues and reduced mitotic figures in the positive control and Budi Shenlian Recipe groups compared to the model group.16S rRNA sequencing showed no significant differences in Chao1 and ACE indices between the high-dose Budi Shenlian Recipe group and the model group.PCoA analysis indicated a distinct microbial community structure between the blank group and mod-el group,with the microbial structure of CRC mice in the Budi Shenlian Recipe group closer to that of the blank group.Compared to the blank group,the model group showed a significant decrease in the proportion of Muribaculaceae,Muribaculum,Alloprevotella,and Prevotellaceae_UCG-001,and a significant increase in Lachnospiraceae_NK4A136_group,Bacteroides,and Helicobacter.After admin-istering Budi Shenlian Recipe to CRC mice,the community structure of some mice partially reverted to the level of the blank group.Transcriptome sequencing revealed that the most significant biological process(BP)among upregulated genes was the negative regulation of macrophage migration,suggesting that Budi Shenlian Recipe can reduce macrophage migration.Moreover,compared to the model group,the proportion of TAMs cells in the tumor tissues of the Budi Shenlian Recipe group significantly decreased(P<0.001).Simultaneously,compared to the model group,the proportion of M1 type TAMs in the Budi Shenlian Recipe group significant-ly increased(P<0.000 1),while the proportion of M2 type TAMs significantly decreased(P<0.05).Immunofluorescence analysis showed the same trend as flow cytometry.The content of TNF-α in tumor tissues of the Budi Shenlian Recipe group significantly in-creased(P<0.001),and IL-10 content significantly decreased(P<0.001).Additionally,compared to the MFMT group,the tumor volume in the BFMT group grew more slowly(P<0.0001).HE staining showed increased necrotic areas,sparser cell arrangement,and reduced pathological mitosis in the BFMT group.Furthermore,compared to the MFMT group,the proportion of TAMs cells in the tumor tissues of the BFMT group significantly decreased(P<0.01).Compared to the MFMT group,the proportion of M1 type TAMs cells in the BFMT group increased(P<0.000 1),while the proportion of M2 type TAMs cells decreased(P<0.01).Immunofluores-cence analysis further confirmed that Budi Shenlian Recipe fecal microbiota transplantation can reduce the proportion of TAMs in the tumor tissues of CRC mice and promote the conversion from M2 to M1 type,thereby reducing immune suppression in the tumor micro-environment.CONCLUSION Budi Shenlian Recipe can improve gut microbiota dysbiosis in CRC mice and exert anti-CRC effects by reducing tumor infiltration of TAMs and modulating the phenotypic transition of TAMs.There may be a certain correlation between these two effects.

Objective To construct microscale rectangular hydrogel grooves and to investigate the morphology and alignment of human umbilical vein endothelial cells(HUVECs)under spatial constraints.Vascular endothelial cell morphology and alignment are important factors in vascular development and the maintenance of homeostasis.Methods A 4-arm polyethylene glycol-acrylate(PEG-acrylate)hydrogel was used to fabricate rectangular microgrooves of the widths of 60 μm,100 μm,and 140 μm.The sizes and the fibronectin(FN)adhesion of these hydrogel microgrooves were measured.HUVECs were seeded onto the FN-coated microgrooves,while the flat surface without micropatterns was used as the control.After 48 hours of incubation,the morphology and orientation of the cells were examined.The cytoskeleton was labelled with phalloidine and the orientation of the cytoskeleton in the hydrogel microgrooves was observed by laser confocal microscopy.Results The hydrogel microgrooves constructed exhibited uniform and well-defined morphology,a complete structure,and clear edges,with the width deviation being less than 3.5%.The depth differences between the hydrogel microgrooves of different widths were small and the FN adhesion is uniform,providing a micro-patterned growth interface for cells.In the control group,the cells were arranged haphazardly in random orientations and the cell orientation angle was(46.9±1.8)°.In contrast,the cell orientation angle in the hydrogel microgrooves was significantly reduced(P<0.001).However,the cell orientation angles increased with the increase in hydrogel microgroove width.For the 60 μm,100 μm,and 140 μm hydrogel microgrooves,the cell orientation angles were(16.4±2.8)°,(24.5±3.2)°,and(30.3±3.5)°,respectively.Compared to that of the control group(35.7%),the number of cells with orientation angles<30° increased significantly in the hydrogel microgrooves of different widths(P<0.001).However,as the width of the hydrogel microgrooves increased,the number of cells with orientation angles<30° gradually decreased(79.9%,62.3%,54.7%,respectively),while the number of cells with orientation angles between 60°-90° increased(P<0.001).The cell bodies in the microgrooves were smaller and more rounded in shape.The cells were aligned along the direction of the microgrooves and corresponding changes occurred in the arrangement of the cell cytoskeleton.In the control group,cytoskeletal filaments were aligned in random directions,presenting an orientation angle of(45.5±3.7)°.Cytoskeletal filaments were distributed evenly within various orientation angles.However,in the 60 μm,100 μm,and 140 μm hydrogel microgrooves,the orientation angles of the cytoskeletal filaments were significantly decreased,measuring(14.4±3.1)°,(24.7±3.5)°,and(31.9±3.3)°,respectively.The number of cytoskeletal filaments with orientation angles<30° significantly increased in hydrogel microgrooves of different widths(P<0.001).However,as the width of the hydrogel microgrooves increased,the number of cytoskeletal filaments with orientation angles<30° gradually decreased,while the number of cytoskeletal filaments with orientation angles between 60°-90° gradually increased(P<0.001).Conclusion Hydrogel microgrooves can regulate the morphology and orientation of HUVECs and mimic to a certain extent the in vivo microenvironment of vascular endothelial cells,providing an experimental model that bears better resemblance to human physiology for the study of the unique physiological functions of vascular endothelial cells.Nonetheless,the molecular mechanism of spatial constraints on the morphology and the assembly of vascular endothelial cell needs to be further investigated.