ABSTRACT:Objective To observe the effect of curcumin on RAW264.7 macrophages induced with LPS and IFNγ(M1)and the mechanisms involved.Methods Curcumin of different concentrations (6.25 μmol/L,12.5μmol/L and 25 μmol/L)was used to treat RAW264.7 macrophages induced with LPS and IFNγ(M1)for 12 h,and RAW264.7 macrophages induced with LPS and IFNγ(M1)were incubated with 20μmol/L GW9662 and 25 μmol/L curcumin for 12 h.Using Real-time PCR,ELISA and Western blotting analysis,we examined the expressions of IL-1β,IL-6,PPARγand phenotype markers M2 (KLF4,FIZZ1,and MGL1 )and the expressions of KLF4 and FIZZ1 when PPARγwas inhibited.Results Curcumin of different concentrations all could inhibit the expressions of IL-1βand IL-6 in RAW264.7 macrophages induced with LPS and IFNγ(M1).Curcumin of different concentra-tions could upregulate the expression of M2 markers (KLF4,FIZZ1 and MGL1)and PPARγin RAW264.7 macro-phages induced with LPS and IFNγ(M1).When M1 macrophages were incubated with curcumin and GW9662,the expression of the M2 phenotype markers was reduced.Conclusion Curcumin polarized the M1 phenotype macro-phages derived from RAW264.7 macrophages to become M2 phenotype through activating PPARγ.

OBJECTIVETo examine the changes in the expression of pro-inflammatory and anti-inflammatory cytokines in the peripheral blood mononuclear cells (PBMCs) and elucidate the inflammatory status of apparently healthy subjects.

METHODSPeripheral blood samples (5 ml) were collected after fasting for more than 8 h from 14 healthy control subjects and 14 apparently healthy subjects with elevated serum high-sensitivity CRP (hsCRP) level (≥ 2.0 mg/L). PBMCs were separated by Ficoll density gradient centrifugation and the total RNA was extracted for real-time quantitative PCR analysis of the inflammatory cytokines, and plasma was separated for ELISA analysis of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expressions.

RESULTSThe gene expressions of TNF-α and MCPIP were significantly increased in PBMCs of apparently healthy subjects, while IL-6 and MCP-1 only showed slight elevations; IL-10 expression in PBMCs decreased significantly in apparently healthy subjects as compared to that in the control group. The results of ELISA showed significantly elevated TNF-α level without significant changes of plasma IL-6 level in apparently healthy subjects.

CONCLUSIONIn apparently healthy subjects with normal lipid levels, chronic low-grade inflammation has occurred shown by elevated expressions of the pro-inflammatory cytokines and lowered expressions of the anti-inflammatory cytokines.

Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Healthy Volunteers ; Humans ; Inflammation ; immunology ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Leukocytes, Mononuclear ; immunology ; Tumor Necrosis Factor-alpha ; blood

Objective To examine the changes in the expression of pro-inflammatory and anti-inflammatory cytokines in the peripheral blood mononuclear cells (PBMCs) and elucidate the inflammatory status of apparently healthy subjects. Methods Peripheral blood samples (5 ml) were collected after fasting for more than 8 h from 14 healthy control subjects and 14 apparently healthy subjects with elevated serum high-sensitivity CRP (hsCRP) level (≥2.0 mg/L). PBMCs were separated by Ficoll density gradient centrifugation and the total RNA was extracted for real-time quantitative PCR analysis of the inflammatory cytokines, and plasma was separated for ELISA analysis of interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α) expressions. Results The gene expressions of TNF-α and MCPIP were significantly increased in PBMCs of apparently healthy subjects, while IL-6 and MCP-1 only showed slight elevations;IL-10 expression in PBMCs decreased significantly in apparently healthy subjects as compared to that in the control group. The results of ELISA showed significantly elevated TNF-α level without significant changes of plasma IL-6 level in apparently healthy subjects. Conclusion In apparently healthy subjects with normal lipid levels, chronic low-grade inflammation has occurred shown by elevated expressions of the pro-inflammatory cytokines and lowered expressions of the anti-inflammatory cytokines.

Objective To examine the changes in the expression of pro-inflammatory and anti-inflammatory cytokines in the peripheral blood mononuclear cells (PBMCs) and elucidate the inflammatory status of apparently healthy subjects. Methods Peripheral blood samples (5 ml) were collected after fasting for more than 8 h from 14 healthy control subjects and 14 apparently healthy subjects with elevated serum high-sensitivity CRP (hsCRP) level (≥2.0 mg/L). PBMCs were separated by Ficoll density gradient centrifugation and the total RNA was extracted for real-time quantitative PCR analysis of the inflammatory cytokines, and plasma was separated for ELISA analysis of interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α) expressions. Results The gene expressions of TNF-α and MCPIP were significantly increased in PBMCs of apparently healthy subjects, while IL-6 and MCP-1 only showed slight elevations;IL-10 expression in PBMCs decreased significantly in apparently healthy subjects as compared to that in the control group. The results of ELISA showed significantly elevated TNF-α level without significant changes of plasma IL-6 level in apparently healthy subjects. Conclusion In apparently healthy subjects with normal lipid levels, chronic low-grade inflammation has occurred shown by elevated expressions of the pro-inflammatory cytokines and lowered expressions of the anti-inflammatory cytokines.

Objective:To investigate the feasibility, effectiveness and safety of ultrasound-guided transperineal prostate biopsy (TPB) with coaxial needle technique, and to improve the pain perception of TPB patients by reducing the number of direct perineal needling.Methods:A total of 200 patients who underwent ultrasound-guided TPB at the first clinical college of Three Gorges University & Yichang Central People′s Hospital from January 2019 to December 2020 were randomly divided into coaxial needle group (coaxial needle positioning puncture, n=100) and traditional group (traditional puncture frame guided repeated transperineal puncture, n=100). Visual analog scale (VAS) was used to evaluate the pain of patients during puncture. The number of samples, time-consuming of puncture, cancer detection rate, VAS pain score and complications between the two groups were compared. Results:The success rate of puncture in the coaxial needle group and the traditional group was 100%, and there was no significant difference in the cancer detection rate between the two groups (48% vs 40%, P>0.05). The average number of samples in the coaxial needle group was larger than that in the traditional group, the average puncture time in the coaxial needle group was less than that in the traditional group, and the average intraoperative VAS score of the coaxial needle group was lower than that of the traditional group, the differences were statistically significant[(14.8±1.8) vs (12.1±1.1), (12.9±1.3)min vs (16.5±1.9)min, (2.6±1.2) vs (4.4±1.4); all P<0.001]. The complication rate of the coaxial needle group was lower than that of the traditional group, the difference was statistically significant (18% vs 39%, P<0.001), the incidences of perineal hematoma and perineal pain in the coaxial needle group were lower than that in the traditional group (1% vs 8%, 8% vs 19%; all P<0.05). Conclusions:Coaxial needle technology for ultrasound-guided TPB can ensure the number of samples and accurate sampling in different areas, significantly reduce the number of direct perineal puncture, improve the pain in the process of puncture, reduce the incidence of postoperative perineal pain, with shorter operation time and fewer complications, which is worthy of clinical promotion.

Objective:To investigate the characteristics of cervical microbiota in patients with HPV (Human Papillomavirus) infection, and to analyze the associations of cervical microbiota and HPV infection or cervicitis.Methods:300 samples underwent HPV nucleic acid testing was collected in this case-control study from June 2019 to April 2020 in the Zhujiang Hospital of Southern Medical University, there were 150 cases allocated in HPV infection group (HPV+), and 150 cases of negative nucleic acid test were non-infectious Group (HPV-). Next-generation sequencing was used to sequence the V4 region of the bacterial 16S rRNA gene, and QIIME pipeline was used to analysis the microbiota composition of the two groups. Wilcoxon rank sum test and Kruskal-Wallis test were used to statistically analysis the differences of the microbiota between groups; and the α diversity and β diversity of the flora between groups were statistically analyzed by Adonis multivariate analysis of variance and Wilcoxon rank sum test.Results:A total of 300 samples were analyzed in this study, of which 150 samples were HPV-positive and 150 samples were HPV-negative; among HPV-positive cases, 132 were infected by high-risk HPV (88.0%), and 18 were low-risk HPV infections (12.0%). The composition of the cervical microbiota were significantly different between the HPV+group and the HPV-group, which in the HPV+group, the α diversity of the cervical microbiota were significantly increased (Shannon index, W=8 174, P<0.000 1; PD whole tree, W=8 887, P=0.001 7). The β diversity of the two groups was significantly different (Binary Jaccard, F=2.325 4, P=0.042 0; Bray Curtis, F=2.136 44, P=0.044 0). The relative abundance of Lactobacillus spp. and L.iners in the HPV+group sample decreased significantly (W=7 730, P<0.000 1; W=8 979, P=0.002 5), accompanied by enriched Achromobacter, Stenotrophomonas, Methylobacterium, Sneathia and Dialister. There was no significant difference in the composition of the cervical microbiota between high-risk HPV infection and low-risk HPV infection ( F=4.100 4, P>0.05). In addition, cervicitis is significantly related to HPV infection (χ2=19.78, P<0.000 1), the composition of cervical flora has similarity features in cervicitis and HPV infection samples. Compared with the normal group, the cervical microbiota of cervicitis with HPV infection is mainly enriched in Achromobacter, Aerococcaceae, Streptococcus, Fusobacteria, and Xanthomonadaceae.Conclusion:The cervical microbiota of patients with HPV infection has a significant dysbiosis, with increased diversity and significant depletion of lactobacillus, accompanied by an increase in the abundance of pathogenic bacteria such as Achromobacter.

Objective:To investigate the characteristics of cervical microbiota in patients with HPV (Human Papillomavirus) infection, and to analyze the associations of cervical microbiota and HPV infection or cervicitis.Methods:300 samples underwent HPV nucleic acid testing was collected in this case-control study from June 2019 to April 2020 in the Zhujiang Hospital of Southern Medical University, there were 150 cases allocated in HPV infection group (HPV+), and 150 cases of negative nucleic acid test were non-infectious Group (HPV-). Next-generation sequencing was used to sequence the V4 region of the bacterial 16S rRNA gene, and QIIME pipeline was used to analysis the microbiota composition of the two groups. Wilcoxon rank sum test and Kruskal-Wallis test were used to statistically analysis the differences of the microbiota between groups; and the α diversity and β diversity of the flora between groups were statistically analyzed by Adonis multivariate analysis of variance and Wilcoxon rank sum test.Results:A total of 300 samples were analyzed in this study, of which 150 samples were HPV-positive and 150 samples were HPV-negative; among HPV-positive cases, 132 were infected by high-risk HPV (88.0%), and 18 were low-risk HPV infections (12.0%). The composition of the cervical microbiota were significantly different between the HPV+group and the HPV-group, which in the HPV+group, the α diversity of the cervical microbiota were significantly increased (Shannon index, W=8 174, P<0.000 1; PD whole tree, W=8 887, P=0.001 7). The β diversity of the two groups was significantly different (Binary Jaccard, F=2.325 4, P=0.042 0; Bray Curtis, F=2.136 44, P=0.044 0). The relative abundance of Lactobacillus spp. and L.iners in the HPV+group sample decreased significantly (W=7 730, P<0.000 1; W=8 979, P=0.002 5), accompanied by enriched Achromobacter, Stenotrophomonas, Methylobacterium, Sneathia and Dialister. There was no significant difference in the composition of the cervical microbiota between high-risk HPV infection and low-risk HPV infection ( F=4.100 4, P>0.05). In addition, cervicitis is significantly related to HPV infection (χ2=19.78, P<0.000 1), the composition of cervical flora has similarity features in cervicitis and HPV infection samples. Compared with the normal group, the cervical microbiota of cervicitis with HPV infection is mainly enriched in Achromobacter, Aerococcaceae, Streptococcus, Fusobacteria, and Xanthomonadaceae.Conclusion:The cervical microbiota of patients with HPV infection has a significant dysbiosis, with increased diversity and significant depletion of lactobacillus, accompanied by an increase in the abundance of pathogenic bacteria such as Achromobacter.

OBJECTIVETo investigate the pathologies of aortic root atherosclerotic lesion in uremic apoE-/- mice and explore the effect of serum from patients with chronic renal insufficiency (CRI) and the uremic toxin, indoxyl sulfate (IS), on the expression of cholesterol transporting receptors and lipid accumulation in macrophages in vitro.

METHODSThe uremic apoE-/- mouse model was established by surgical operation. Frozen sections of the aortic root were collected from uremic apoE-/- mice, sham-operated apoE-/- mice and C57BL/6J mice and stained with oil red O to calculate the relative area of atherosclerotic plaque. Murine macrophage RAW264.7 cell line was treated for 12 h with different concentrations of IS or serum samples from CRI patients and healthy individuals, and the mRNA expressions of cholesterol transporting receptors (SR-A1, CD36, ABCA1, ABCG1 and SR-B1) were detected. After treatment for 24 h, the cells were induced into foam cells to determine lipid contents using oil red O staining.

RESULTSThe relative area of the atherosclerotic plaques in the aortic root increased significantly in uremic apoE-/- mice compared with that in sham-operated apoE-/- mice. CRI serum (5%) and IS (250 µmol/L) obviously increased the mRNA expression of CD36 and lipid accumulation in the macrophages, but did not affect the mRNA expression of other cholesterol transporting receptors.

CONCLUSIONCRI can accelerate the progression of atherosclerosis through the mechanism that IS in CRI serum promotes lipid accumulation in macrophages by enhancing the mRNA expression of CD36, which contributes to the formation of foam cells.

Animals ; Apolipoproteins E ; Cell Line ; Foam Cells ; chemistry ; Humans ; Indican ; pharmacology ; Lipids ; chemistry ; Macrophages ; chemistry ; Mice ; Mice, Inbred C57BL ; Plaque, Atherosclerotic ; pathology ; Renal Insufficiency, Chronic ; blood

Objective To investigate the pathologies of aortic root atherosclerotic lesion in uremic apoE-/-mice and explore the effect of serum from patients with chronic renal insufficiency (CRI) and the uremic toxin, indoxyl sulfate (IS), on the expression of cholesterol transporting receptors and lipid accumulation in macrophages in vitro. Methods The uremic apoE-/-mouse model was established by surgical operation. Frozen sections of the aortic root were collected from uremic apoE-/-mice, sham-operated apoE-/- mice and C57BL/6J mice and stained with oil red O to calculate the relative area of atherosclerotic plaque. Murine macrophage RAW264.7 cell line was treated for 12 h with different concentrations of IS or serum samples from CRI patients and healthy individuals, and the mRNA expressions of cholesterol transporting receptors (SR-A1, CD36, ABCA1, ABCG1 and SR-B1) were detected. After treatment for 24 h, the cells were induced into foam cells to determine lipid contents using oil red O staining. Results The relative area of the atherosclerotic plaques in the aortic root increased significantly in uremic apoE-/- mice compared with that in sham-operated apoE-/- mice. CRI serum (5%) and IS (250 μmol/L) obviously increased the mRNA expression of CD36 and lipid accumulation in the macrophages, but did not affect the mRNA expression of other cholesterol transporting receptors. Conclusion CRI can accelerate the progression of atherosclerosis through the mechanism that IS in CRI serum promotes lipid accumulation in macrophages by enhancing the mRNA expression of CD36, which contributes to the formation of foam cells.

Objective To investigate the pathologies of aortic root atherosclerotic lesion in uremic apoE-/-mice and explore the effect of serum from patients with chronic renal insufficiency (CRI) and the uremic toxin, indoxyl sulfate (IS), on the expression of cholesterol transporting receptors and lipid accumulation in macrophages in vitro. Methods The uremic apoE-/-mouse model was established by surgical operation. Frozen sections of the aortic root were collected from uremic apoE-/-mice, sham-operated apoE-/- mice and C57BL/6J mice and stained with oil red O to calculate the relative area of atherosclerotic plaque. Murine macrophage RAW264.7 cell line was treated for 12 h with different concentrations of IS or serum samples from CRI patients and healthy individuals, and the mRNA expressions of cholesterol transporting receptors (SR-A1, CD36, ABCA1, ABCG1 and SR-B1) were detected. After treatment for 24 h, the cells were induced into foam cells to determine lipid contents using oil red O staining. Results The relative area of the atherosclerotic plaques in the aortic root increased significantly in uremic apoE-/- mice compared with that in sham-operated apoE-/- mice. CRI serum (5%) and IS (250 μmol/L) obviously increased the mRNA expression of CD36 and lipid accumulation in the macrophages, but did not affect the mRNA expression of other cholesterol transporting receptors. Conclusion CRI can accelerate the progression of atherosclerosis through the mechanism that IS in CRI serum promotes lipid accumulation in macrophages by enhancing the mRNA expression of CD36, which contributes to the formation of foam cells.