Research progress on association between macrophages and ischemia-reperfusion injury / 器官移植

Ischemia-reperfusion injury (IRI) is an extremely complicated pathophysiological process, which may occur during the process of myocardial infarction, stroke, organ transplantation and temporary interruption of blood flow during surgery, etc. As key molecules of immune system, macrophages play a vital role in the pathogenesis of IRI. M1 macrophages are pro-inflammatory cells and participate in the elimination of pathogens. M2 macrophages exert anti-inflammatory effect and participate in tissue repair and remodeling and extracellular matrix remodeling. The balance between macrophage phenotypes is of significance for the outcome and treatment of IRI. This article reviewed the role of macrophages in IRI, including the balance between M1/M2 macrophage phenotype, the mechanism of infiltration and recruitment into different ischemic tissues. In addition, the potential therapeutic strategies of targeting macrophages during IRI were also discussed, aiming to provide reference for alleviating IRI and promoting tissue repair.

Collagen content and C-X-C motif chemokine ligand 12 expression in neoplastic breast stroma

SUMMARY OBJECTIVE: This study aimed to evaluate the expression of C-X-C motif chemokine ligand 12 and its C-X-C chemokine receptor type 4, and the tumor-stroma ratio using collagen stromal content of breast cancer samples, correlating it with clinicopathological data. METHODS: Through a retrospective cohort study, samples were obtained from female patients, over 18 years of age, with the disease in stages 1-4, who underwent mastectomy or lumpectomy. The biopsies were provided by the Oncology sector of the Hospital das Clínicas of Universidade Federal de Pernambuco, Recife city, in 2011-2014, including samples of invasive ductal carcinoma, ductal carcinoma in situ, or benign changes (fibroadenoma and hypertrophy), which were analyzed between 2020 and 2022 by immunohistochemistry for the expression of stromal cell characteristics. Collagen content was tested by Gomori staining and digital analysis of images. RESULTS: Absence of stromal expression of C-X-C motif chemokine ligand 12 was associated with longer disease-free survival (disease-free survival=0.481), and expression of C-X-C chemokine receptor type 4 was associated with lower disease-free survival. An association was observed between clinicopathological variables and stromal expression of chemokines, that is, an association of stromal C-X-C motif chemokine ligand 12 with histological grade, angiolymphatic invasion, and an association between C-X-C chemokine receptor type 4 expression and histological grade. Analyses of digital pixels images of collagen and tumor cells showed a lower percentage of collagen in the invasive ductal carcinoma samples (39%), unlike samples without neoplasms (78%). CONCLUSION: Low expression of C-X-C motif chemokine ligand 12 may be associated with a worse prognosis for breast cancer. Collagen staining analyzed using digital images represents an opportunity for clinical application and is indicative of the prognosis of the tumor microenvironment in breast carcinoma.

Effect of siRNA interference with C-C chemokine ligand 5 on biological characteristics of laryngeal carcinoma cells / 中国生物制品学杂志

@#Objective To investigate the expression of C-C chemokine ligand 5(CCL5) in head and neck squamous cell carcinoma(HNSCC),and explore the effect of CCL5 on the biological characteristics of laryngeal carcinoma cells.Methods Gene Expression Profiling Interactive Analysis(GEPIA) database was used to investigate the expression of CCL5 in HNSCC.The laryngeal carcinoma cells TU177 were transfected with siRNA(siRNA group),and the control(NC) group was set up.The cell proliferation,migration,cycle and apoptosis of each group were detected by CCK8 assay,cell scratch test and flow cytometry respectively.RT-PCR and Western blot were used to detect the knock-down efficiency of CCL5 and the mRNA transcription and protein expression of multidrug resistance protein 2(MRP2) and bcl-2-associated x protein(Bax).Results The expression of CCL5 in HNSCC was higher than that in normal tissues(P <0.05).Compared with NC group,siRNA showed higher knock-down efficiency(t=12.898 and 22.656 respectively,each P <0.01);siRNA interference with CCL5 inhibited the proliferation and migration of laryngeal carcinoma cells,and promoted the late apoptosis of laryngeal carcinoma cells and the expression of apoptosis protein Bax(t=2.600～11.667,each P <0.05).Conclusion CCL5 was highly expressed in HNSCC,while siRNA interference with CCL5 inhibited the proliferation,migration and promoted apoptosis of laryngeal carcinoma cells TU177 by up-regulating the expression of Bax,which laid a foundation of the possibility of CCL5 as a new target for the treatment of laryngeal carcinoma.

Prognostic value of CC motif chemokine receptor 2 in the microenvironment of glioblastoma and its relationship with the immune cell infiltration / 肿瘤研究与临床

Objective:To explore the prognostic biomarkers of glioblastoma (GBM) in the tumor microenvironment (TME) and its function.Methods:A total of 169 GBM samples of 161 GBM patients were collected from the Cancer Genome Atlas (TCGA) database. ESTIMATE algorithm in R4.1.0 software was used to calculate the proportion of immune components and stromal components in TME, which were expressed as immune score and stromal score, respectively. According to the median value of the two scores, 169 GBM samples were divided into the high score group and the low score group, respectively, 84 each in each group (those whose scores were equal to the median were not involved in the grouping). The differentially expressed genes (DEG) [false discovery rate (FDR) < 0.05] between the high score group and the low score group of the two scores were obtained by using limma package, and the co-up-regulated and co-down-regulated DEG of the two scores were obtained by using Venn program. Based on the STRING database, the protein interaction (PPI) network of co-up-regulated and down-regulated DEG of immune score and stromal score was constructed, and the top 30 genes with connectivity were selected. Univariate Cox proportional hazard model analysis of overall survival (OS) of 161 GBM patients in the TCGA database was performed on co-up-regulated and down-regulated DEG between immune score and stromal score by using R4.1.0 software to obtain the DEG affecting OS. The intersection of the DEG obtained from PPI analysis and Cox analysis was taken as the prognostic core genes. According to the median expression value of prognostic core genes in GBM samples from the TCGA database, 161 patients were divided into prognostic core genes high expression group and low expression group (patients whose scores were equal to the median were not involved in the grouping), with 80 cases in each group. Kaplan-Meier survival analysis of OS was performed by using R4.1.0 software. GSEA 4.2.1 software was used to perform gene set enrichment analysis (GSEA) on all genes with transcriptome data of GBM patients in the two groups of the TCGA databases, and the main enriched functions of the two groups of genes were obtained. The CIBERSORT algorithm was used to test the accuracy of the proportion of tumor infiltrating immune cell (TIC) subsets in 169 GBM samples from the TCGA database, and 57 GBM samples were finally obtained. Immune cells with differential expression levels and immune cells related to the expression of prognostic core genes among the samples with different expression levels of prognostic core genes were analyzed; Venn program was used to obtain the intersection of immune cells with differential levels and related immune cells, and differentially expressed TIC related to expressions of prognostic core genes in GBM were obtained.Results:Based on the immune score and stromal score of GBM samples in the TCGA database, a total of 693 co-up-regulated and co-down-regulated DEG of both scores were screened out. After the intersection of 78 DEG related to OS obtained by univariate Cox regression analysis and 30 DEG obtained by PPI network results, CC motif chemokine receptor 2 (CCR2) was identified as the prognostic core gene ( HR = 1.294, 95% CI 1.060-1.579, P = 0.011). GBM patients with CCR2 high expression had worse OS compared with those with CCR2 low expression ( P = 0.009). GSEA analysis showed that genes in the CCR2 high expression group were mainly enriched in immune-related pathways, while genes in the CCR2 low expression group were mainly enriched in metabolism-related pathways. Among 57 screened GBM samples, there were differences in the levels of 3 immune cells between the CCR2 high expression group and the CCR2 low expression group ( P < 0.05). CCR2 expression was correlated with the levels of 9 immune cells (all P < 0.05). Venn program analysis showed that differentially expressed 3 TIC in GBM related to CCR2 gene expression were obtained; among them, M2 macrophages were positively correlated with CCR2 expression, while T follicular helper cell and activated NK cells were negatively correlated with CCR2 expression. Conclusions:CCR2 may be the core gene related to the prognosis in the TME of GBM. As reference, the level of CCR2 can help to predict the status of TME and prognosis in GBM patients, which is expected to provide a new direction for the treatment of GBM.

Preliminary study on the ability of 68Ga-Pentixafor PET/CT to differentiate between adrenal aldosterone-producing adenoma and nonfunctional adenoma / 中华内科杂志

Objective:To evaluate the ability of 68Ga-Pentixafor (nuclide ligand imaging agents for chemokine receptor 4) PET/CT to differentiate between aldosterone-producing adenoma (APA) and adrenal nonfunctional adenoma (NFA), and to assess how well this imaging method correlates with clinical features and postoperative outcomes. Methods:This was a cross-sectional study involving 73 APA and 12 NFA patients who received 68Ga-Pentixafor PET/CT imaging at Peking Union Medical College Hospital from August 2018 to October 2021. The receiver operating characteristic (ROC) curve was used to evaluate the differential value of visual analysis and the maximum standard uptake value (SUV max) of the focus on APA and NFA. The related factors of SUV max, and its predictive effect on postoperative outcomes were analyzed using Pearson or Spearman analysis and χ2 text. Results:68Ga-Pentixafor PET/CT imaging was positive in 64 APA patients (sensitivity=87.7%) and negative in all 12 NFA patients (specificity=100%). The area under the ROC curve with SUV max differentiating APA and NFA was 0.932 ( P<0.001). When the SUV max cut-off point was 6.23, the sensitivity was 80.8% and the specificity was 100%. The SUV max correlated positively with lesion size ( r=0.598) and aldosterone/renin activity ratio ( r=0.313) and correlated negatively with potassium level ( r=-0.286), renin activity ( r=-0.240) and age of diagnosis ( r=-0.273) (all P<0.05). Of the patients who underwent adrenalectomy and received more than 6 months of post-surgical follow-up, the clinical complete remission rate was higher for 68Ga-Pentixafor PET/CT imaging-positive patients than imaging-negative patients (24/39 vs. 0/4, P=0.031). Conclusions:68Ga-Pentixafor PET/CT is effective at differentiating between APA and NFA. The SUV max of 68Ga-Pentixafor PET/CT correlates with age at onset, lesion size, and the severity of clinical manifestations, and is able to predict postoperative outcomes.

Research progress of vitamin D, monocyte chemoattractant protein-1 and Hashimoto's thyroiditis / 中国医师杂志

Hashimoto thyroiditis (HT) is one of the most common autoimmune thyroid disease, and its pathogenesis has not been fully clarified at present. Most people believe that it is induced by mental stress, overwork, infection, stress, environmental pollution, unreasonable diet structure (such as high iodine diet) and other factors on the basis of genetic defects and genetic susceptibility. Vitamin D is a steroid hormone that maintains the balance of calcium and phosphorus metabolism in the body, regulating bone and mineral salt metabolism. Monocyte chemoattractant protein-1 (MCP-1) is a member of the chemokine CC family. It binds to chemokine receptor (CCR) and participates in immune inflammatory response. In recent years, more and more studies have found that vitamin D and MCP-1 are involved in the occurrence and development of many immune diseases, including Hashimoto thyroiditis. This article reviews the new research progress of the role of vitamin D and MCP-1 in Hashimoto thyroiditis.

Effects of Hcy and lncRNA ANRIL on atherosclerotic inflammatory response / 中国医师杂志

Objective:To study the relationship between the level of serum homocysteine (Hcy) and the antisense non coding gene (ANRIL) of long chain non coding RNA (lncRNA) cell cycle dependent kinase inhibitor 2B gene, and the effect on Atherosclerosis inflammation, that is, the expression of interleukin-10 (IL-10) and Monocyte chemoattractant protein-1 (MCP-1) in Human umbilical vein endothelial cell (HUVEC).Methods:HUVEC was cultured in vitro and cells were treated with different concentration gradients (blank control group, 0.5, 1.0, 2.0, 5.0 mmol/L) of Hcy. The expression level of lncRNA ANRIL was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of MCP-1 and IL-10. LipoFilter transfection reagents were used to transfect shANRIL and shNC into different cells, respectively. In the above experiment, the optimal Hcy concentration (5.0 mmol/L) was selected for intervention for 24 hours. Western blot was used to detect the protein expression levels of MCP-1 and IL-10.Results:After 24 hours of intervention with different concentrations of Hcy in HUVEC, Hcy significantly damaged endothelial cells, and the higher the Hcy concentration, the more severe the cell damage. Compared with the blank control group, the Hcy intervention group showed an increase in lncRNA ANRIL and MCP-1, while IL-10 decreased (all P<0.05); As the concentration of Hcy intervention increases, IL-10 decreases, while lncRNA ANRIL and MCP-1 increased (all P<0.05). Compared with the blank control group, the Hcy group, the shNC+ Hcy group, and the shANRIL+ Hcy group had lower levels of IL-10 protein expression and higher levels of MCP-1 protein expression (all P<0.05). Compared with the shANRIL+ Hcy group, the Hcy group and the shNC+ Hcy group had lower levels of IL-10 protein expression and higher levels of MCP-1 protein expression (all P<0.05). There was no statistically significant difference in the expression levels of IL-10 protein and MCP-1 protein between the shNC+ Hcy group and the Hcy group (all P>0.05). Conclusions:Hcy upregulates MCP-1 expression and downregulates IL-10 expression by promoting lncRNA ANRIL expression. Thus, it can promote cellular inflammatory reaction and participate in Atherosclerosis.

The serum levels of FCGBP, SAA1, CXCL10 in children with mycoplasma pneumoniae pneumonia and their relationship with prognosis / 中国医师杂志

Objective:To investigate the serum levels and clinical significance of Fc fragment of the IgG-binding protein (FCGBP), serum amyloid protein A1 (SAA1), and CXC chemokine ligand 10 (CXCL10) in children with mycoplasma pneumoniae pneumonia (MPP) and their relationship with prognosis.Methods:A prospective study was conducted on 122 children with MPP admitted to the department of pediatrics of the 970th Hospital of the Joint Logistics Support Force of the Chinese People′s Liberation Army from January 2019 to December 2021. According to the severity and prognosis of MPP, they were divided into mild and severe groups, good prognosis group, and poor prognosis group. Forty healthy children who underwent physical examination during the same period were set as the control group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum levels of FCGBP, SAA1, and CXCL10 in each subject, and to compare the differences in serum levels of FCGBP, SAA1, and CXCL10 among different groups. Multivariate logistic regression analysis was used to investigate the influencing factors of poor prognosis in MPP patients. The diagnostic value of individual and combined detection of serum procalcitonin (PCT), FCGBP, SAA1, and CXCL10 for poor prognosis in MPP children by analyzing the receiver operating characteristic (ROC) curve.Results:The levels of serum FCGBP [(115.68±10.57)ng/ml, (78.41±6.73)ng/ml, (12.55±3.25)ng/ml], SAA1 [(34.18±3.72)mg/L, (25.54±2.63)mg/L, (6.74±0.82)mg/L], and CXCL10 [(714.26±55.64)ng/L, (353.74±42.67)ng/L, (106.25±12.92)ng/L] in the severe MPP group were significant higher than those in the mild MPP group and the control group, with statistical significance (all P<0.05). The white blood cell (WBC), neutrophil percentage, C reactive protein (CRP), erythrocyte sedimentation rate (ESR), PCT, lactate dehydrogenase (LDH), D-dimer (D-D), FCGBP, SAA1, CXCL10 of the children in the poor prognosis group were significantly higher than those in the good prognosis group, and the differences were statistically significant (all P<0.05). Multivariate logistic regression analysis showed that increased PCT ( OR=1.603, 95% CI: 1.190-2.160), FCGBP ( OR=1.757, 95% CI: 1.115-2.770), SAA1 ( OR=1.900, 95% CI: 1.327-2.720) and CXCL10 ( OR=1.704, 95% CI: 1.212-2.397) were independent risk factors for poor prognosis of MPP children (all P<0.05). The combined detection of serum PCT, FCGBP, SAA1, and CXCL10 had a significantly higher diagnostic value for the risk of poor prognosis in children with MPP than a single indicator. Conclusions:The elevated levels of serum FCGBP, SAA1, and CXCL10 in children with MPP are associated with the severity of MPP and are independent risk factors for poor prognosis in MPP patients.

Role of serum CX3CR1 in the diagnosis of coronary artery stenosis and in the evaluation of prognosis after percutaneous coronary intervention / 中国基层医药

Objective:To investigate the role of serum CX3CR1 in the diagnosis of coronary artery stenosis and in the evaluation of prognosis after percutaneous coronary intervention.Methods:A total of 101 patients with coronary artery stenosis (≥ 50% stenosis) confirmed with coronary angiography (CAG) in Haiyang People's Hospital from January 2018 to May 2019 who were followed up till May 2021 were included in the observation group. Thirty-four healthy individuals who underwent physical examination during the same period were included in the control group. Patients in the observation group were divided into an in-stent restenosis group (ISR group, n = 28) and a non-ISR group ( n = 73). The expression of CX3CR1 was detected. The incidence of adverse cardiac events was calculated. The sensitivity, specificity, and area under the curve (AUC) plotted for the use of CX3CR1 to diagnose coronary artery stenosis and predict adverse cardiac events were evaluated. Results:The expression of CX3CR1 in the observation group was (3.95 ± 1.05) μg/L, which was significantly higher than (2.30 ± 0.65) μg/L in the control group ( t = 2.87, P < 0.05). The receiver operating characteristic curve analysis showed that the AUC, sensitivity, and specificity of the use of CX3CR1 in diagnosing coronary artery stenosis were 0.892, 75.2%, and 88.2%. The incidence of non-fatal myocardial infarction, angina pectoris, heart failure, and cardiac death in the ISR group was significantly higher compared with the non-ISR group ( χ2 = 8.06, 7.17, 8.06, 7.17, all P < 0.05). The receiver operating characteristic curve analysis results showed that the AUC value of CX3CR1 in predicting non-fatal myocardial infarction, angina pectoris, heart failure, and cardiac death were 0.786, 0.895, 0.997, and 0.887, respectively. Conclusion:CX3CR1 is highly expressed in coronary artery stenosis, which can provide a reference for the diagnosis and prognostic evaluation of coronary artery stenosis.

Effects of thoracic segment epidural anesthesia on inflammatory factors in patients undergoing lung cancer surgery / 中国基层医药

Objective:To investigate the effects of thoracic segment epidural anesthesia on inflammatory factors in patients undergoing lung cancer surgery.Methods:The clinical data of 136 patients who underwent lung cancer surgery in the Second People's Hospital of Liaocheng from June 2020 to May 2022 were retrospectively analyzed. According to anesthesia methods, these patients were divided into an observation group ( n = 89) and a control group ( n = 47). The observation group was given thoracic segment epidural anesthesia, while the control group was given remifentanil infusion anesthesia. The tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-10 (IL-10) levels in the epithelial lining fluid collected from the non-dependent lung, the plasma levels of TNF-α, IL-6, and malondialdehyde, arterial partial pressure of oxygen/inhaled oxygen fraction, the incidence of complications, the incidence of re-operations, numeric rating scale score, and the length of hospital stay were compared between the two groups. The effects of different anesthesia methods on lung cancer surgery were evaluated. Results:In each group, TNF-α, IL-6, and IL-10 levels in the epithelial lining fluid were significantly increased 30 minutes after termination of one-lung ventilation (T2) compared with those measured before one-lung ventilation (T1) ( t = 7.71, 77.10, 7.59, 3.41, 57.51, 5.74, all P < 0.05). In the observation group, TNF- α [(1.59 ± 0.53) ng/L, (1.89 ± 0.64) ng/L] measured at T1 and T2, IL-6 [(2.96 ± 0.82) ng/L] and IL-10 [(1.99 ± 0.53) ng/L] measured at T1 were significantly higher compared with those measured at the corresponding time points in the control group ( t = 10.45, 2.59, 2.00, 7.19, all P < 0.05). In the observation group, IL-6 measured at T2 [(38.91 ± 5.84) ng/L] was significantly lower than that in the control group ( t = 33.25, P < 0.001), and IL-10 measured at T2 [(2.51 ± 0.67) ng/L] was slightly, but not significantly higher than that in the control group ( P > 0.05). There was no significant difference in the plasma level of TNF- α measured at T1 and T2 between the two groups (both P > 0.05). Plasma levels of IL-6 in the two groups [(42.98 ± 5.29) ng/L, (27.93 ± 4.17) ng/L] measured at T2 were significantly increased compared with those measured at T1 ( t = 54.14, 61.06, both P < 0.001). In the observation group, TNF-α measured at T2 [(1.60 ± 0.56) ng/L] and IL-6 measured at T1 and T2 [(0.92 ± 0.16) ng/L, (27.93 ± 4.17) ng/L] were significantly lower compared with the control group ( t = 3.39, 6.96, 18.20, all P < 0.05). There were no significant differences in plasma level of malondialdehyde, arterial partial pressure of oxygen/inhaled oxygen fraction, numeric rating scale score, the incidence of complications, the incidence of re-operation, and the length of hospital stay between the two groups (all P > 0.05). Conclusion:Thoracic segment epidural anesthesia can reduce the local inflammatory response of the lung during lung cancer surgery.

Aconite aqueous extract inhibits the growth of hepatocellular carcinoma through CCL2-dependent enhancement of natural killer cell infiltration / 中西医结合学报

OBJECTIVE@#Aconite is a traditional Chinese herbal medicine that has been found to inhibit the development of liver cancer; however, its exact molecular mechanisms in this process remain unclear. This study explores how aconite aqueous extract (AAE) inhibits hepatocellular carcinoma (HCC).@*METHODS@#An in vivo mouse model of subcutaneous liver cancer was established. After AAE treatment, immunohistochemistry (IHC) was used to determine the effect of AAE on natural killer (NK) cells. Subsequently, C57BL/6 mice were used to establish the subcutaneous tumor model, and a group of these mice were treated with anti-PK163 antibody to remove NK cells, which was verified by flow cytometry and IHC. The effect of AAE on the proliferation of HCC cells in vitro was determined using cell counting kit-8. The effect of AAE on chemokine production in HCC cells was measured using real-time quantitative polymerase chain reaction and an enzyme-linked immunosorbent assay. The effect of AAE on the migration of NK cells was determined using a transwell assay. Finally, the molecular mechanism was investigated using the Western blotting method.@*RESULTS@#We demonstrated that the ability of AAE to induce overexpression of the cytokine C-C motif chemokine ligand 2 (CCL2) in HCC cells is fundamental to the infiltration of NK cells into the tumor bed. Mechanistically, we found that the upregulation of CCL2 was achieved by the activation of c-Jun N-terminal kinase but not extracellular regulated protein kinase or p38.@*CONCLUSION@#Our findings suggest that AAE can be used as an effective immune adjuvant to enhance antitumor immunity by increasing NK cell infiltration into tumors, which could help to improve the efficacy of HCC treatments. Please cite this article as: Yang KD, Zhang X, Shao MC, Wang LN. Aconite aqueous extract inhibits the growth of hepatocellular carcinoma through CCL2-dependent enhancement of natural killer cell infiltration. J Integr Med. 2023; 21(6): 575-583.

Bone marrow mesenchymal stem cells exosome miR-126-3p targets CCR1 to inhibit the malignant proliferation and metastasis of non-small cell lung cancer cells / 西安交通大学学报(医学版)

【Objective】 To investigate the effects of miR-126-3p targeting chemokine receptor 1 (CCR1) in exosomes derived from bone marrow mesenchymal stem cells (BMSC) on the proliferation, migration, and invasion of lung cancer cells. 【Methods】 BMSC cells were cultured; exosomes were extracted and identified by the exosomal marker proteins CD63 and TSG101. After exosome culture of A549 cells for different durations (0, 24, 48, and 72 h), cell survival rate was detected by CCK-8, mRNA levels of miR-126-3p and CCR1 were detected by qRT-PCR, and cell migration and invasion abilities were detected by Transwell assay. The relative expressions of CCR1, epithelial cadherin (E-cad), neural cadherin (N-cadherin), and Vimentin were detected by Western blotting. 【Results】 Exosomes had round or oval cup-shaped structures with bright edges and dark middle, with a particle size distribution of about 152 nm, expressing CD63 and TSG101 proteins. The expression of miR-126-3p in exosomes was higher than that in A549 cells. The expression of miR-126-3p was low in A549 cells and that of CCR1 mRNA was high. However, after co-culture with exosomes, the expression of miR-126-3p in A549 cells was increased, while the expression of CCR1 was decreased. A549 cells were cocultured with exosomes for 0, 24, 48, and 72 h. The survival rate, migration and invasion abilities, CCR1 gene and protein expression levels, and N-cad and Vimentin protein expression levels of A549 cells decreased gradually with the extension of culture time. The level of miR-126-3p and the expression of E-cad protein increased gradually with the extension of culture time. 【Conclusion】 The co-culture of exosomes derived from bone marrow mesenchymal stem cells with A549 cells can increase the expression level of miR-126-3p, and miR-126-3p can reduce the proliferation, migration, and invasion of A549 cells by targeting the inhibition of CCR1 expression.

Scientific basis of acupuncture on mesenchymal stem cells for treating ischemic stroke / 中国针灸

The scientific basis of acupuncture on mesenchymal stem cells (MSCs) for treating ischemic stroke (IS) is discussed. MSCs transplantation has great potential for the treatment of tissue damage caused by early stage inflammatory cascade reactions of IS, but its actual transformation is limited by various factors. How to improve the homing efficiency of MSCs is the primary issue to enhance its efficacy. As such, the possible mechanisms of acupuncture and MSCs transplantation in inhibiting inflammatory cascade reactions induced by IS are explored by reviewing literature, and a hypothesis that acupuncture could promote the secretion of stromal cell-derived factor-1α (SDF-1α) from ischemic foci to regulate SDF-1α/CXC chemokine receptor 4 (CXCR4) axis, thereby improving the homing efficiency of MSCs transplantation, exerting its neuroprotective function, and improving the bed transformation ability, is proposed.

Regulatory Mechanism of Mangiferin Combined with Bortezomib on Malignant Biological Behavior of Burkitt Lymphoma and Its Effect on Expression of CXC Chemokine Receptors / 中国实验血液学杂志

OBJECTIVE@#To analyze the effects of mangiferin combined with bortezomib on the proliferation, invasion, apoptosis and autophagy of human Burkitt lymphoma Raji cells, as well as the expression of CXC chemokine receptors (CXCRs) family, and explore the molecular mechanism between them to provide scientific basis for basic research and clinical work of Burkitt lymphoma.@*METHODS@#Raji cells were intervened with different concentrations of mangiferin and bortezomib alone or in combination, then cell proliferation was detected by CCK-8 assay, cell invasion ability was detected by Transwell chamber method, cell apoptosis was detected by Annexin V/PI double-staining flow cytometry, apoptosis, autophagy and Akt/mTOR pathway protein expression were detected by Western blot, and the expression changes of CXCR family was detected by real-time quantitative PCR (RT-qPCR).@*RESULTS@#Different concentrations of mangiferin intervened Raji cells for different time could inhibit cell viability in a concentration- and time-dependent manner (r =-0.682, r =-0.836). When Raji cells were intervened by combination of mangiferin and bortezomib, compared with single drug group, the proliferation and invasion abilities were significantly decreased, while the apoptosis level was significantly increased (P <0.01). Mangiferin combined with bortezomib could significantly up-regulate the expression of pro-apoptotic protein Bax and down-regulate the expression of anti-apoptotic protein Bcl-2 after intervention in Raji cells. Caspase-3 was also hydrolyzed and activated, and then induced the apoptosis of Raji cells. Mangiferin combined with bortezomib could up-regulate the expression of LC3Ⅱ protein in Raji cells, and the ratio of LC3Ⅱ/LC3Ⅰ in cells was significantly up-regulated compared with single drug or control group (P <0.01). Mangiferin combined with bortezomib could significantly inhibit the phosphorylation levels of Akt and mTOR, inhibit the proliferation and invasion of Raji cells by inhibiting Akt/mTOR pathway, and induce cell autophagy and apoptosis. Mangiferin and bortezomib could down-regulate the expressions of CXCR4 and CXCR7 mRNA after single-agent intervention in Raji cells, and the down-regulations of CXCR4 and CXCR7 mRNA expression were more significant when the two drugs were combined (P <0.01). Mangiferin alone or combined with bortezomib had no significant effect on CXCR5 mRNA expression in Raji cells (P >0.05), while the combination of the two drugs could down-regulate the expression of CXCR3 (P <0.05).@*CONCLUSION@#Mangiferin combined with bortezomib can synergistically inhibit the proliferation and invasion of Raji cells, and induce autophagy and apoptosis. The mechanism may be related to the inhibition of Akt/mTOR signaling pathway, down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bax, and the inhibition of the expression of CXCR family.

Relationship between expression of chemokine receptor 2 and basic fibroblast growth factor in aqueous humor and prognosis of trabeculectomy in patients with acute primary angle-closure glaucoma / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the relationship between the levels of chemokine receptor 2(CXCR2)and basic fibroblast growth factor(bFGF)in aqueous humor and the prognosis of trabeculectomy in patients with acute primary angle-closure glaucoma(APACG).METHODS: A total of 80 cases(80 eyes)APACG patients who underwent trabeculectomy in our hospital from June 2020 to January 2022 were collected in the case group. According to the postoperative efficacy, they were grouped into a success group of 60 cases(60 eyes)and a failure group of 20 cases(20 eyes). Another 86 cataract patients(86 eyes)who underwent phacoemulsification with normal intraocular pressure in our hospital during the same period were included in the control group. Enzyme linked immunosorbent assay was applied to detect the levels of CXCR2 and bFGF in aqueous humor. ROC curve was applied to analyze the value of predicting trabeculectomy failure in APACG patients by the levels of CXCR2 and bFGF in aqueous humor. Furthermore, multivariate Logistic regression was applied to analyze the influencing factors of trabeculectomy failure in APACG patients.RESULTS: The levels of CXCR2 and bFGF in the aqueous humor of the case group were significantly higher than those of the control group(P&#x003C;0.05). The levels of CXCR2 and bFGF in the aqueous humor of the failed group and the proportion of patients with postoperative shallow anterior chamber were significantly higher than those of the successful group(P&#x003C;0.05). The AUC for predicting trabeculectomy failure in APACG patients using CXCR2 and bFGF levels alone and in combination was 0.885, 0.883 and 0.953, respectively. CXCR2 and bFGF were independent risk factors for trabeculectomy failure in APACG patients(P&#x003C;0.05).CONCLUSION: The levels of CXCR2 and bFGF in the aqueous humor of APACG patients are obviously elevated, and both are risk factors for trabeculectomy failure.

Bushen Jianpi Jiedu Liyan Formula Regulates Homing-associated Cell Adhesion Molecules in Rat Model of IgA Nephropathy / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Bushen Jianpi Jiedu Liyan formula on the expression of integrin alpha 4 beta 1 （α4 β1）， vascular cell adhesion molecule-1 （VCAM-1）， stromal-derived factor-1 （SDF-1）， and chemokine receptor-4 （CXCR4） in the small intestine and bone marrow of the rat model of immunoglobulin A（IgA） nephropathy. MethodA total of 120 male SD rats were used to establish the IgA nephropathy model by intragastric administration of bovine serum albumin （BSA）， subcutaneous injection of CCl4， and tail vein injection of lipopolysaccharide （LPS）. The successfully modeled rats were randomized into blank， model， lotensin （63 mg·kg-1）， and low-， medium-， and high-dose （10.4， 20.81， 41.62 g·kg-1， respectively） Bushen Jianpi Jiedu Liyan formula groups （n=16）. The rats were treated with corresponding drugs according to their body weight. After 7 weeks of administration， the rats were sacrificed for the collection of samples， and the protein and mRNA levels of α4 β1， VCAM-1， SDF-1， and CXCR4 in the small intestine and bone marrow were determined by immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction， respectively. ResultCompared with the blank group， the model group showed increased red blood cell count in the urine at the 10th， 12th， 14th， 16th weeks （P<0.01）， and such increases were reduced in the drug intervention groups （P<0.05）， especially in the medium-dose Bushen Jianpi Jiedu Liyan formula group （P<0.05）. Compared with those in the blank group， the protein levels of α4 β1， VCAM-1， SDF-1， and CXCR4 in the intestinal lamina propria in the model group were up-regulated （P<0.05）， and such un-regulations were inhibited in the drug intervention groups （P<0.05）. Compared with the model group， medium-dose Bushen Jianpi Jiedu Liyan formula down-regulated the protein levels of SDF-1 and CXCR4 in the intestinal lamina propria （P<0.05）. Compared with the blank group， the model group showed down-regulated mRNA levels of α4 β1 and SDF-1 and up-regulated mRNA levels of VCAM-1 and CXCR4 （P<0.05）. Compared with the model group， the drug intervention groups showed down-regulated mRNA levels of SDF-1 and CXCR4 （P<0.05）. ConclusionBushen Jianpi Jiedu Liyan formula regulates the expression of α4 β1， VCAM-1， SDF-1， and CXCR4 in the intestinal lamina propria to inhibit the homing effect of plasma cells， which may be associated with the Toll-like receptor-mediated activation of immune response. Bushen Jianpi Jiedu Liyan formula can down-regulate the expression of adhesion molecules to inhibit the proliferation of plasmocytes in circulation， so as to reduce the renal injury of IgA nephropathy.

Modified Erchentang Alleviates Inflammation in Chronic Obstructive Pulmonary Disease via Midkine/Notch2/Hey1 Signaling Pathway in Rats / 中国实验方剂学杂志

ObjectiveTo investigate the molecular mechanism of the anti-inflammatory effect of Erchentang in the lung tissue of the rat model of chronic obstructive pulmonary disease （COPD） via the heparin-binding factor （Midkine）/transmembrane receptor protein （Notch2）/Hey1 signaling pathway. MethodSixty SD rats were randomized into normal group， model group， modified Erchentang （5， 10， 20 g·kg-1·d-1） groups， and Notch1 pathway inhibitor （γ-secretase inhibitor， DAPT， 0.02 g·kg-1） group， with 10 rats in each group. The rat model of COPD was established by cigarette smoke combined with lipopolysaccharide （LPS）. After the modeling， the rats were administrated with corresponding drugs by gavage， and those in the normal and model groups were administrated with normal saline by gavage for 21 days. The levels of Midkine， cytokine-induced neutrophil chemoattractant-1 （CINC-1）， macrophage-derived chemokine （MDC）， chemokine ligand 5 （CXCL5）， neutrophil elastase （NE）， and nuclear factor-kappa B （NF-κB） p65 in bronchoalveolar lavage fluid （BALF） were determined by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and immunohistochemistry were respectively employed to determine the mRNA and protein levels of Midkine， Notch2， and Hey1 in the lung tissue. ResultCompared with the normal group， the modeling increased the levels of Midkine， CINC-1， MDC， CXCL5， NE， and NF-κB p65 in BALF （P<0.01） and up-regulated the mRNA and protein levels of Midkine， Notch2， and Hey1 in the lung tissue （P<0.01）. Compared with the model group， medium- and high-dose modified Erchentang and DAPT lowered the levels of Midkine， CINC-1， MDC， CXCL5， and NF-κB p65 in BALF （P<0.01） and down-regulated the mRNA levels of Midkine， Notch2， and Hey1 （P<0.01）. ConclusionModified Erchentang may inhibit the inflammation in COPD rats by down-regulating the expression of Midkine， Notch2， and Hey1 and reducing the content of Midkine， CINC-1， MDC， and CXCL5.

Construction of interferon alpha/beta receptor subunit 1 gene knockout Caco-2 cell line based on CRISPR/Cas9 system / 中国生物制品学杂志

@#Objective To knockout interferon alpha/beta receptor subunit 1(IFNAR1) gene in human colorectal adenocarcinoma cells Caco-2 using clustered regularly interspaced short palinmic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system to construct IFNAR1 knockout Caco-2 cell line.Methods The single guide RNA(sgRNA)sequence was designed to specifically recognize the exon region of IFNAR1 gene using CRISPR/Cas9 technology,and the LentiCRISPRv2-IFNAR1-sgRNA recombinant plasmid was constructed.Caco-2 cells were infected with the plasmid packaged by lentivirus and screened by puromycin resistance.The obtained monoclonal cell lines were cultured by limited dilution method,which were verified for the effect of IFNAR1 gene knockout by target gene sequencing and Western blot,and detected for the mRNA levels of CXC chemokine ligand 10(CXCL10)and interferon-stimulatd gene 20(ISG20)in IFNAR1knockout cells by adding exogenous IFNβ.Results Sequencing results of plasmid LentiCRISPRv2-IFNAR1-sgRNA showed that the insertion sites were all located at the sticky end of BsmBⅠenzyme digestion.Two IFNAR1 knockout monoclonal cell lines were obtained.The sequencing results showed that Caco-2-IFNAR1-KO1 had 5 bp deletion in the sixth exon of IFNAR1,and Caco-2-IFNAR1-KO2 had 18 bp deletion and 1 bp insertion in the seventh exon.Compared with wild-type Caco-2 cells,Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells showed no expression of IFNAR1 protein.Compared with no IFNβ stimulation,the mRNA levels of CXCL10 gene(t = 0.566 and 1.268 respectively,P>0.05)and ISG20 gene(t =1.522 and 1.733 respectively,P>0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells stimulated by 50 ng/mL IFNβ showed no significant increase.While compared with those of wild-type Caco-2 cells,the mRNA levels of CXCL10gene(t = 6.763 and 6.777 respectively,P<0.05)and ISG20 gene(t = 5.664 and 5.65 respectively,P<0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells decreased significantly under the stimulation of 50 ng/mL exogenous IFNβ.Conclusion Caco-2 cell line with IFNAR1 knockout was successfully constructed by using CRISPR/Cas9 technology,and the downstream molecules activated by IFNAR(interferon alpha/beta receptor)in this cell line were obviously inhibited,which provided a powerful tool for further exploration of the innate immune response and replication packaging mechanism of Caco-2 cells after virus infection.

Advances in the Study of Chemokine-like Factor Superfamily Members in Tumors / 中国肺癌杂志

Chemokine-like factor-like MARVEL transmembrane domain containing member/chemokine-like factor superfamily member (CMTM/CKLFSF) including CKLF and CMTM1-CMTM8 are a new family of proteins linking chemokines and transmembrane superfamilies. CMTM not only have broad chemotactic activities, but also associate with hematopoietic system, immune system, and tumor development and metastasis closely. CMTM proteins are involved in key biological processes of cancer development, which include activation and recycling of growth factor receptors, cell proliferation and metastasis, and regulation of the tumor immune microenvironment. This is a new focus of research on the relationship between CMTM and tumors, because CMTM4/CMTM6 can be considered as a regulator for programmed cell death ligand 1 (PD-L1). This paper reviews the role of CMTM family members on cancer, especially in tumor growth, metastasis and immune escape, summarize the latest findings on the relationship between CMTM and non-small cell lung cancer, and explores the potential clinical value of CMTM as a novel drug target or biomarker.
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Expression Profile and Clinical Significance of Cytokines and Chemokines in Patients with Epstein-Barr Virus-Associated Hemophagocytic Lymphohistiocytosis / 中国实验血液学杂志

OBJECTIVE@#To investigate the cytokine/chemokine profile in patients with Epstein-Barr virus (EBV)-related hemophagocytic lymphohistiocytosis (HLH), and assess the prognostic value of survival.@*METHODS@#Serum levels of thirty-eight cytokines/chemokines were measured by multiple cytokine assay kit in EBV-related HLH patients, EBV-infected patients, and controls. The expression profile of cytokines/chemokines was compared among groups. The changes of cytokine/chemokine expression in active and remission stage of EBV-related HLH patients were also compared, and the prognostic values for survival were evaluated.@*RESULTS@#Serum levels of interferon-α2 (IFN-α2), interleukin (IL)-6, and IL-7 in EBV-related HLH patients were 33.67(23.23-68.78) pg/ml, (74.95±25.53) pg/ml, and 35.35(19.50-63.55) pg/ml, respectively, which were significantly higher than those in EBV-infected patients［IFN-α2: 16.07(9.87-29.63); IL-6: 55.91±20.29; IL-7: 20.40(13.35-31.40)］ and controls ［IFN-α2: 11.02(4.67-21.25); IL-6:42.64±13.41; IL-7: 16.95(14.95-33.78)］(all P<0.05). Serum levels of IL-8, IL-9, and marcophage-derived chemokine (MDC) in EBV-related HLH patients were 11.00(7.50-15.27) pg/ml, 81.30(40.79-111.0) pg/ml, and (512.6±128.7) pg/ml, respectively, which were significantly higher than those in controls ［IL-8: 6.80(5.56-8.38); IL-9: 41.30(29.82-67.91); MDC: 384.1±156.6］(all P<0.05), but there was no remarkable differences compared with EBV-infected patients (P>0.05). Serum IFN-α2, IL-6, IL-7, IL-8, IL-9, and MDC in survival and death groups of EBV-related HLH patients were analyzed by receiver operating characteristic curve with area under curve of 0.781, 0.778, 0.633, 0.805, 0.562, and 0.657, respectively (P=0.019, 0.021, 0.269, 0.015, 0.607, and 0.190). IFN-α2, IL-6, and IL-8 had good predictive effect on survival. Serum level of IFN-α2, IL-6, and MDC of EBV-related HLH patients in remission stage were significantly lower than those in active stage (P<0.05), while IL-7, IL-8, and IL-9 were not different (P>0.05).@*CONCLUSION@#IFN-α2, IL-6, IL-7, IL-8, IL-9, and MDC may take part in the pathogenesis of EBV-related HLH.