Immature stroma and high infiltration of CD15+ cells are predictive markers of poor prognosis in different subsets of patients with pancreatic cancer.

Preoperative treatment is commonly carried out for borderline resectable pancreatic ductal adenocarcinoma (PDAC). However, the relationship between the combination of immune cells in the tumor microenvironment and their intratumoral heterogeneity along with their association with histological findings remains unclear, especially in patients receiving preoperative chemotherapy. We aimed to explore the therapeutic strategies for patients with PDAC with poor prognosis after receiving chemotherapy based on histological and immunological microenvironmental classifications. We investigated the correlation between the prognosis and histological immune microenvironmental factors of patients who initially underwent surgery (n = 100) and were receiving gemcitabine plus nab-paclitaxel (GEM + nabPTX) as preoperative chemotherapy (n = 103). Immune profiles were generated based on immune cell infiltration into the tumor, and their correlation with patient outcomes and histological features was analyzed. Tumor-infiltrating neutrophils (TINs) were identified as independent poor prognostic factors using multivariate analysis in both surgery-first and preoperative chemotherapy groups. The patients were further classified into four groups based on immune cell infiltration into the tumor. Patients with high CD15 infiltration into the tumor and immature stroma at the cancer margins showed the worst prognosis in the preoperative chemotherapy group. The analysis of mRNA expression and immunohistochemical features revealed that CXCR2, the receptor for CXCL8, was correlated with disease-free and overall survival. We inferred that patients with immature stroma at the margins and high infiltration of CD15+ neutrophils within the tumor showed the worst prognosis and they could particularly benefit from treatment with inhibitors targeting CXCR2 or CXCL8.

Clinical Significance Of Baseline And Post-Treatment CXCR1 Expression In Women With Breast Cancer Having Received Neoadjuvant Chemo Therapy.

Objectives: To examine the chemokine receptor type 1 expression in breast cancer tissues before and after neoadjuvant chemotherapy, and its relationship with pathological response to neoadjuvant chemotherapy and other clinical variables. Method: The prospective study was conducted at Kafrelsheikh University Hospital, Egypt, from November 2018 to March 2021, and comprised female patients with new histopathologically proven breast cancer eligible for chemotherapy. Paraffin blocks of tumourspecimens were stained immunohistochemically using concentrated rabbit anti-human chemokine receptor type 1 polyclonal antibody kits. The patients were followed up for treatment response, disease recurrence and mortality. Data was analysed using SPSS 25. RESULTS: Of the 100 patients with mean age 50.2±12.1 years, 40(40%) in group A with mean age 55.1±9.3 showed marked response and 60(60%) in group B with mean age 47.0±12.7 yearsshowed mild/moderate response (p<0.001). Group A patients had significantly lower baseline and post-treatment chemokine receptor type 1 expression compared to group B patients (p<0.05). The change in chemokine receptor type 1 expression was not significantly different (p>0.05). Patients with tumour grade 3 had significantly higher baseline chemokine receptor type 1 expression compared to patients with tumour grade 2. Tumourstage and post-treatment chemokine receptor type 1 expression were also significantly interlinked (p<0.05). Multivariate regression analysisidentified patients'age, baseline chemokine receptor type 1 and post-treatment chemokine receptor type 1 expressions as predictors of treatment response. CONCLUSIONS: There was found to be an association between baseline and post-treatment chemokine receptor type 1 expression in breast cancer tissues and pathological response to neoadjuvant chemo therapy in such 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.

Relationship between neutrophil chemotactic function and chemokine receptor in the early stage of deep second- and third-degree burns / 中国基层医药

Objective:To investigate the relationship between neutrophil chemotactic function and chemokine receptor in the early stage of deep second- and third-degree burns.Methods:Twenty patients with severe burns (burn group) who received treatment within 6 hours after burns in Yantai Yeda Hospital from January 2019 to June 2020 were included in this study. Twenty healthy controls (healthy group) who concurrently received physical examination in the same hospital were also included. The general data and laboratory examination indexes in each group were analyzed. The correlation between neutrophil chemotactic function and chemokine receptor was evaluated.Results:There were no significant differences in general data between the two groups (all P > 0.05). At 1, 3 and 5 days after admission, the number of neutrophils, the number of white blood cells, and procalcitonin, C-reactive protein, interleukin-6, interleukin-10, and tumor necrosis factor-α levels in the burn group were significantly higher than those in the healthy control groups ( F = 12.56, 13.45, 15.78, 17.83, 22.56, 13.39, 10.82, all P < 0.05). At 1, 3 and 5 days after admission, neutrophil migration distance in the burn group was (1 510.22 ± 108.45) μm, (1 380.90 ± 115.67) μm, (1 026.10 ± 95.48) μm, respectively, which were significantly shorter than (1 944.67 ± 139.20) μM in the healthy control group ( t = 23.44, 25.67, 27.52, all P < 0.05). At 5 days after admission, chemokine receptors 1 and 2 positive rates in the burn group were (47.40 ± 1.76)% and (75.33 ± 2.42)%, respectively, which were significantly lower than (95.24 ± 4.89)% and (97.78 ± 2.10)% in the healthy control groups ( t = 4.92, 5.67, both P < 0.05). Correlation analysis showed that neutrophil migration distance was positively correlated with chemokine receptor expression in patients with deep second- and third-degree burns ( r = 0.72, 0.61, both P < 0.05). Conclusion:Neutrophil chemotactic function and chemokine receptor expression decrease in the early stage of deep second- and third-degree burns.

Diagnosis and differential diagnosis value of immune marker CCR7 in chronic lymphocytic leukemia and other chronic B-cell lymphoproliferative disorders / 中华检验医学杂志

Objective:To investigate the clinical significance of cc-chemokine receptor 7 (CCR7) as a potential diagnostic or differential marker for chronic lymphocytic leukemia (CLL).Methods:A total number of 643 patients with B-cell chronic lymphoproliferative diseases (B-CLPD) admitted to the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2018 were enrolled. The patients included 327 cases of CLL, 58 cases of mantle cell lymphoma (MCL), 34 cases of follicular lymphoma (FL), 36 cases of marginal zone lymphoma (MZL), 10 cases of hair-cell leukemia or its variants (HCL/HCLV-v), 40 cases of Waldorf′s macroglobulinemia (WM), 48 cases of CD5 +B-cell chronic lymphoproliferative disease unclassified (B-CLPD-U) and 90 cases of CD5 -B-CLPD-U. At the same time, 20 samples from healthy people from the medical examination center of our hospital were used as normal controls. Flow cytometry was used to detect the immune-phenotype and CCR7 expression level in B-CLPD patients, and Fluorescence in situ hybridization (FISH) was used to analyze the genomic alterations: the ataxia telangiectasia mutant gene (ATM) deletion, the 13q14 deletion, the P53 deletion and trisomy 12. Sanger sequencing was used to analyze gene mutations of splicing factor 3B subunit 1 (SF3B1), NOTCH1, tumor protein 53 (TP53) and immunoglobulin heavy chain variable region (IGHV). Measurement data were compared by Mann-Whitney test, and the positive rates were compared by chi-square test. The diagnostic value and optimal positive cutoff value of CCR7 were calculated using receiver operating characteristic (ROC) curve. Results:The positive rates of CCR7 expression in typical CLL and atypical CLL were 90.8% (257/283) and 84.1% (37/44), respectively, and there was no significant difference of the positive rates (χ 2=1.228, P=0.268) between groups. The positive expression rates of CCR7 in CLL, MCL, CD5 +B-CLPD-U, CD5 -B-CLPD-U, FL, WM, HCL/HCL-v and MZL were 89.9% (294/327), 10.3% (6/58), 6.3% (3/48), 8.9% (8/90), 0, 0, 0 and 13.9% (5/36) respectively, and the median mean fluorescence intensity (MFI) was 278 (246, 307), 114 (106, 128), 112 (106, 117), 110 (104, 121), 108 (105, 119), 111 (105, 124), 112 (108, 115) and 109 (105, 120) respectively. Compared with CLL, the positive expression rates of CCR7 in other types of B-CLPDs were lower significantly (χ 2=181.3, 177.8, 232, 164.7, 180.8, 62.6, 129, P<0.01). In addition, the sensitivity, specificity and accuracy of CCR7 for distinguishing CLL from other types of B-CLPD were 89.9%, 93.0% and 92.3%, respectively. The positive expression rate of CD49d in CCR7 +CLL patients was 13.9%, which was significantly lower than that in CCR7 -CLL patients (42.1%) (χ 2=7.6, P=0.01). The coincidence rate of 13q14 deletion was 50.3% in CCR7 +CLL patients, which was significantly higher than that in CCR7 -CLL patients (20%) (χ 2=6.56, P=0.01). Conclusions:The CC-chemokine receptor 7 (CCR7) antigen is an effective marker for the diagnosis and identification of chronic lymphocytic leukemia (CLL). The expression level of CCR7 in clinical specimens can distinguish CLL from other pathological subtypes of B-CLPDs.

[Early changes of chemotactic function of peripheral blood neutrophil of patients with severe burns and the influence factor].

Objective: To observe the early changes of chemotactic function of peripheral blood neutrophil of patients with severe burns and the influence factor. Methods: Seven severe burn patients who met the inclusion criteria and were admitted to Suzhou Hospital Affiliated to Nanjing Medical University in 6 hours post burns from January to May 2019 were selected and included in burn group (4 males and 3 females, aged (36±10) years). Seven healthy volunteers with normal physical examination results in the Physical Examination Center of the same hospital in the same period of time were included in healthy control group (5 males and 2 females, aged (35±8) years). A prospective and controlled study was performed. (1) The venous blood of 2 mL was taken from each patient in burn group on post admission day (PAD) 1, 3, 5 and venous blood of 2 mL was taken from each volunteer in healthy control group for routine detection of white blood cell count, platelet count, neutrophil count, serum procalcitonin level, and C-reactive protein level. (2) The venous blood of patients and healthy volunteers was taken as before for measuring interleukin-6 (IL-6), IL-10, and tumor necrosis factor α (TNF-α) by enzyme-linked immunosorbent assay. (3) The venous blood of patients and healthy volunteers was taken as before, and peripheral blood neutrophils were isolated by Ficoll density gradient centrifugation. The chemotactic distance of neutrophil was detected by agarose chemotaxis test, and the positive expression rates of chemokine receptor CXCR1 and CXCR2 of patients in burn group on PAD 3 and volunteers in healthy control group were detected by flow cytometer. Data were statistically analysed with analysis of variance for repeated measurement, t test, and Bonferroni correction. Results: (1) The platelet count of patients in burn group on PAD 1, 3, 5 was close to that of volunteers in healthy control group respectively (t=0.55, 0.44, 0.12, P>0.05). The counts of neutrophil and white blood cell and the expression levels of serum procalcitonin and C-reactive protein of patients in burn group on PAD 1, 3, 5 were significantly higher than those of volunteers in healthy control group (t=196.96, 273.31, 45.22, 3.46, 4.18, 5.55, 4.36, 5.26, 11.13, 64.94, 89.97, 84.31, P<0.01). (2) The level of IL-6 of patients in burn group on PAD 1, 3, 5 was significantly higher than that of volunteers in healthy control group respectively (t=187.43, 213.54, 195.74, P<0.01), the level of IL-10 of patients in burn group on PAD 1, 3, 5 was significantly higher than that of volunteers in healthy control group respectively (t=21.47, 11.13, 6.23, P<0.01), and the level of TNF-α of patients in burn group on PAD 1, 3, 5 was significantly higher than that of volunteers in healthy control group respectively (t=5.27, 7.89, 15.58, P<0.01). (3) The chemotactic distances of neutrophil of patients in burn group were (1 479±102), (1 395±82), and (1 017±91) µm respectively on PAD 1, 3, 5, which were significantly shorter than (1 902±120) µm of volunteers in healthy control group (t=7.11, 9.23, 15.55, P<0.01). (4) The CXCR1 and CXCR2 positive expression rates of neutrophil of patients in burn group on PAD 3 were (48.3±1.6)% and (79.0±1.8)%, respectively, which were significantly lower than (95.4±4.5)% and (97.8±2.1)% of volunteers in healthy control group (t=27.13, 23.10, P<0.01). Conclusions: The chemotactic dysfunction of peripheral blood neutrophil was detected in the early stage of severe burn patients, which may be related to the decreases of CXCR1 and CXCR2.

Effective Treatments for Bladder Cancer Affecting CXCL9/CXCL10/CXCL11/CXCR3 Axis: A Review.

Bladder cancer (BC) originates mainly from the epithelial compartment of the bladder, which is defined as transitional cell carcinoma or urothelial cell carcinoma. About 70% of patients with BC will survive five years from diagnosis. Previous studies revealed that the immune system and its mediators, particularly chemokines, play a crucial role in modulating responses against BC. Chemokines, which serve as chemoattractants for leukocytes, are small proteins that can initiate inflammatory and anti-inflammatory immune responses and also are associated with many aspects of both regulation and progression of mentioned responses. Additionally, these immune mediators can interfere with the other tumor-related processes, including tumor proliferation, neovascularization, and metastases. Among these chemokines, CXC chemokines, including CXCL9, CXCL10, and CXCL11, are recognized as the main ligands of C-X-C motif chemokine receptor 3 (CXCR3) and contribute to related immune responses after therapeutic strategies for BC. Evidence suggests that the production of these chemokines can have two important implications. First, these mediators can trigger the accumulation of CD8+ T cells that can contribute to the elimination of the tumor. Secondly, the production of these chemokines by tumor tissue may trigger the migration and activation of immune cells including myeloid-derived suppressor cells and regulatory T cells, which act in favor of the tumor and its progress. Therefore, in this review, we describe the latest therapeutic approaches based on targeting this axis's components and subsequent immune phenomenon.

Correlation analysis between CXC chemokine receptor 6 and prognosis of renal clear cell carcinoma / 肿瘤研究与临床

Objective@#To investigate the correlation between CXC chemokine receptor 6 (CXCR6) and prognosis of renal clear cell carcinoma.@*Methods@#A total of 100 patients with renal clear cell carcinoma who underwent surgery in the First People's Hospital of Ziyang from January 2013 to October 2014 were selected. Immunohistochemistry was used to detect the expression of CXCR6 in 100 cases of renal clear cell carcinoma and adjacent normal tissues. The patients were followed up to observe the positive rate of CXCR6 expression and the factors affecting overall survival (OS) and progression-free survival (PFS) of renal clear cell carcinoma and adjacent normal tissues.@*Results@#The positive rate of CXCR6 expression in renal clear cell carcinoma tissues was higher than that in adjacent tissues, and the difference was statistically significant [46.0% (46/100) vs. 20.0% (20/100), χ2 = 15.287, P < 0.01]. The positive expression rate of CXCR6 in patients with clinical stage Ⅲ-Ⅳ, lymph node metastasis and pathological grade Ⅲ-Ⅳ was higher than that in patients with clinical stage Ⅰ- Ⅱ, pathological grade Ⅰ- Ⅱ and without lymph node metastasis, and the difference was statistically significant (all P < 0.05). A total of 92 patients were followed up and 40 died. The follow-up time reached to (35-60) months and the median follow-up time was 48.9 months. Kaplan-Meier and log-rank test showed that patients with positive CXCR6 expression had lower 3-year OS and PFS rate compared with patients with negative CXCR6 expression, and the difference was statistically significant (3-year OS rate: 52.1% vs. 78.6%, χ2 = 10.027, P = 0.001; 3-year PFS rate: 48.3% vs. 67.8%, χ2 = 4.344, P = 0.037). Cox regression multivariate analysis showed pathological grade (OS: OR = 2.154, 95% CI 1.547-9.517, P = 0.023; PFS: OR = 1.235, 95% CI 1.109-5.917, P = 0.042), lymph node metastasis (OS: OR = 1.412, 95% CI 1.109-5.917, P = 0.041; PFS: OR = 1.841, 95% CI 1.354-8.994, P = 0.010), CXCR6 expression (OS: OR = 1.864, 95% CI 1.358-6.813, P = 0.031; PFS: OR = 1.457, 95% CI 1.127-6.884,P = 0.025) were independent risk factors of OS and PFS for renal clear cell carcinoma patients treated by the surgery.@*Conclusions@#The positive expression of CXCR6 is an independent risk factor for OS and PFS of renal clear cell carcinoma. The prognosis of patients with positive CXCR6 expression is poor.

[Study on the effects of target-silencing CXCR3 expression on malignant proliferation of hepatocellular carcinoma].

Objective: To explicit, the expression of chemokine receptor 3 in HCC tissues and its relationship with overall survival of patients, and to explore the effect of targeted silencing CXCR3 gene on proliferation of hepatocellular carcinoma cells and its mechanism of action. Methods: The expression of CXCR3 in 60 cases of hepatocellular carcinoma and its adjacent tissues were detected by immunohistochemistry. The clinicopathological correlations between the expression levels of CXCR3 in hepatoma tissues of liver cancer patients were analyzed and univariate Kaplan-Meier survival analysis was performed in combination with follow-up data. Huh7 hepatoma cells were infected with lentivirus LV-CXCR3-shRNA. The effects of CXCR3 deletion on proliferation of hepatoma cells were determined by CCK-8 assay and tumor-bearing nude mice experiment. Results: CXCR3 was highly expressed in HCC tissues, and the overall survival rate (OS) of patients with high CXCR3 expression was significantly lower than that of patients with low expression. After the CXCR3 gene was successfully silenced in Huh7 hepatocellular carcinoma cells, the proliferation ability of Huh7 cells was significantly inhibited in vitro, and the tumor growth rate of nude mice was slowed down, and the activity of JAK-STAT pathway in Huh7 cells was decreased, and the levels of c-MYC and Bcl-xl protein were decreased. In addition, deletion of CXCR3 can effectively inhibit IL-6-mediated JAK-STAT pathway activation. Conclusion: CXCR3 high expression indicated that the survival rate was poor, and the target silencing of CXCR3 gene could inhibit the proliferation of hepatocellular carcinoma cells and maybe related to inhibition of JAK-STAT pathway activity. CXCR3 may be a potential target for the treatment of hepatocellular carcinoma.

[Effect of cigarette smoke exposure on the expression of CCR7 and levels of Th1/Th2 cytokines in asthmatic rats].

Objective: To investigate the effect of cigarette smoke exposure on the expression of CC Chemokine receptor 7 (CCR7) and levels of Th1/Th2 cytokines in asthmatic rats. Methods: Forty Wistar rats were randomly divided into four groups: control group, asthma group, smoke exposure group, asthma-smoke exposure group. The asthma group were sensitized with ovalbumin (OVA) and Aluminum hydroxide at day 1, 8 and challenged with OVA at day 15 by atomization for 8 weeks.While control group was sensitized and challenged with normal saline instead of OVA.The smoke exposure group was sensitized and challenged with normal saline instead of OVA followed passive smoking for 8 weeks. The asthma-smoke exposure group was challenged with OVA followed passive smoking. The pathological changes of different groups were observed by HE-staining. CCR7 was semiquantitatively analyzed in lungs by immunohistochemistry.The concentration of CC chemokine ligand (CCL)19, CCL21, interferon (IFN)-γ and interleukin (IL)-4 in peripheral blood and CCL19 and CCL21 in bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent (ELESA) assay. Results: In asthma group, smoke exposure group and asthma-smoke exposure group, the various degrees of inflammatory reaction appeared in lung tissue and the asthma-smoke exposure group was with the most significant reaction. In the lung tissues of the rats from asthma group, smoke exposure group and asthma-smoke exposure group, the average optical density (AOD) of CCR7 were significantly higher than those in control group (0.350±0.023, 0.252±0.022, 0.400±0.029 vs 0.180±0.020, all P<0.01). The AOD of CCR7 of asthma-smoke exposure group was much higher than both that in asthma group and in smoke exposure group (both P<0.01). In asthma group, smoke exposure group and asthma-smoke exposure group, the concentrations of both CCL19 and CCL21 in peripheral blood and BALF were significantly higher than that in control group (all P<0.01). The concentrations of both CCL19 and CCL21 in peripheral blood and BALF of asthma-smoke exposure group were significantly higher than the results in asthma group and in smoke exposure group (all P<0.01). The concentrations of IFN-γ in peripheral blood of asthma group and asthma-smoke exposure group were lower than those in control group [(33±3), (17±3) vs (70±4) pg/ml], but asthma-smoke exposure group was much lower than the results in asthma group (all P<0.01). The concentration of IFN-γ in peripheral blood of smoke exposure group[(100±5)pg/ml]was higher than that in control group and asthma-smoke exposure group (both P<0.01). In asthma group, smoke exposure group, asthma-smoke exposure group, the concentrations of IL-4 in peripheral blood were significantly higher than those in control group [(54±4), (42±4), (76±4) vs (30±4) pg/ml, all P<0.01]. The concentrations of IL-4 in peripheral blood of asthma-smoke exposure group was significantly higher than those in asthma group and in smoke exposure group (both P<0.01). Conclusion: Cigarette smoke could enhance the expression of CCR7 and its ligand, and it can also result in exacerbations of asthma by reducing the expression level of IFN-γ (the representative of Th1 cytokine) and increasing the expression level of IL-4 (the representative of Th2 cytokine).

[Relationship between C-C chemokine receptor type 2 and P38 mitogen-activated protein kinase signaling pathway in the spinal cord of rats with bone cancer pain].

Objective: To investigate the relationship between C-C chemokine receptor type 2(CCR2) and P38 mitogen-activated protein kinase (P38MAPK) signaling pathway in the spinal cord of rats and further clarify the mechanism of bone cancer pain (BCP). Methods: A total of 92 healthy female SD rats, of which 60 were subjected to behavioral tests using a ciliary mechanical stimulation needle. SD rats were randomly divided into six groups: sham operation group (group S), bone cancer pain group (group B), sham operation + DMSO solvent group (group SD), bone cancer pain + DMSO solvent group (group BD), sham operation + RS102895 CCR2 inhibitor group (group SR), bone cancer pain + RS102895 CCR2 inhibitor group (group BR), and Von Frey was used in the behavioral test. Another 32 SD rats were randomly divided into the following 8 groups (n=4): sham operation group (group S), bone cancer pain 5 d group (group B5), bone cancer pain 9 d group (group B9), bone cancer pain 14 d group (group B14), bone cancer pain + DMSO solvent group (group BD), bone cancer pain + RS102895 CCR2 inhibitor 0.5 h group (group BR0.5 h), bone cancer pain + RS102895 CCR2 inhibitor 4 h group (group BR4 h), bone cancer pain + RS102895 CCR2 inhibitor 12 h group (group BR12 h). Western blot was used to detect the expression of P38, p-P38 and CCR2 in spinal cord of rats. Results: At day 5, 7, 9, 14, 21 post-injection, mechanical withdrawal thresholds of group S were(30.9±1.5), (31.9±1.2), (32.0±1.1), (31.6±1.5), (32.2±1.4)g respectively, the mechanical withdrawal thresholds of group B were( 26.4±0.7), (24.4±0.8), (21.4±0.8), (13.5±0.4), (9.9±0.2)g respectively, the mechanical withdrawal thresholds in group B decreased obviously versus group S, and the differences were statistically significant(t=-13.177, -16.660, -23.778, -35.574, -48.401, all P<0.01). At day 9 post-injection, the mechanical withdrawal thresholds in SD, BD, SR and BR groups were (32.4±1.7), (19.4±1.1), (32.1±1.3), (26.3±1.0) g respectively, the difference was statistically significant (F=224.681, P<0.01), and the mechanical withdrawal thresholds in group BD decreased obviously versus group SD, while the mechanical withdrawal thresholds in group BR increased obviously versus group BD. The expression levels of p-P38 in spinal cord of group S, group B5, group B9 and group B14 were(0.08±0.03), (0.20±0.05), (0.40±0.17), (0.65±0.14)respectively, the expression levels of CCR2 were(0.08±0.04), (0.18±0.05), (0.30±0.09), (0.58±0.07)respectively, the difference was statistically significant(F=19.123, 40.746, all P<0.01), and the expression of p-P38 and CCR2 in group B9 were showed a significant up-regulation versus group S. The expression levels of p-P38 in spinal cord of group BD, group BR0.5 h, group BR4 h and group BR12 h were (0.57±0.06), (0.17±0.11), (0.03±0.01), (0.25±0.11)respectively, and the difference was statistically significant(F=29.582, P<0.01). The expression of p-P38 in group BR0.5 h, BR4 h, BR12 h showed a significant down-regulation versus group BD. Conclusion: CCR2 in the spinal cord may be involved in the development of bone cancer pain by activating P38MAPK signaling pathway in rats.

Effects of irbesartan combined with resuvastatin on early diabetic nephropathy complicated with cardiovascular disease and its influence on SDF-1/CXCR4 expression / 中国基层医药

Objective To investigate the effect of irbesartan combined with resuvastatin on early diabetic nephropathy( DN) complicated with cardiovascular disease,and its influence on SDF -1/CXCR4 expression. Methods From January 2016 to December 2017,130 early DN patients complicated with cardiovascular disease in the Second People＇s Hospital of Linhai were selected as study objects,and according to the digital table,they were randomly divided into three groups: irbesartan group(control group 1,n＝43),resuvastatin group(control group 2,n＝43) and irbesartan combined with resuvastatin group (observation group,n＝44).The clinical efficacy,blood pres-sure,blood lipids and SDF -1/CXCR4 expression were compared among the three groups.Results After treatment,the total effective rate in the observation group was 93.18%(41/44 ),which in the control group 1 was 69.77%(30/43),which in the control group 2 was 74.42%(32/43),there was statistically significant difference among the three groups(χ2＝12.341,10.106,all P＜0.05).After treatment,the levels of SBP and DBP in the observation group had statistically significant differences compared with those in the control group 1 and control group 2( F＝23.087,20.249,all P＜0.05).After treatment,there were statistically significant differences in TC,TG,HDL-C and LDL-C between the observation group and control group 1,control group 2(F＝18.408,15.623,14.852,9.845,all P＜0.05).After treatment,the level of SDF -1 in the observation group was significantly higher than that before treatment,and which was also significantly higher than that in the control group 1 and control group 2(F＝21.085,P＜0.05).After treatment,the positive rate of CXCR4 in the observation group was significantly higher than that before treatment,and compared with that in the control group 1 and control group 2,the difference was statistically significant(F＝23.641,P＜0.05).Conclusion Irbesartan combined with resuvastatin in the treatment of early DN patients with cardiovascular disease has significant clinical efficacy,can effectively improve blood pressure and blood lipid levels,significantly improve the level of SDF -1 and CXCR4 positive rate.

Study on the effects of target-silencing CXCR3 expression on malignant proliferation of hepatocellular carcinoma / 中华肝脏病杂志

Objective@#To explicit, the expression of chemokine receptor 3 in HCC tissues and its relationship with overall survival of patients, and to explore the effect of targeted silencing CXCR3 gene on proliferation of hepatocellular carcinoma cells and its mechanism of action.@*Methods@#The expression of CXCR3 in 60 cases of hepatocellular carcinoma and its adjacent tissues were detected by immunohistochemistry. The clinicopathological correlations between the expression levels of CXCR3 in hepatoma tissues of liver cancer patients were analyzed and univariate Kaplan-Meier survival analysis was performed in combination with follow-up data. Huh7 hepatoma cells were infected with lentivirus LV-CXCR3-shRNA. The effects of CXCR3 deletion on proliferation of hepatoma cells were determined by CCK-8 assay and tumor-bearing nude mice experiment.@*Results@#CXCR3 was highly expressed in HCC tissues, and the overall survival rate (OS) of patients with high CXCR3 expression was significantly lower than that of patients with low expression. After the CXCR3 gene was successfully silenced in Huh7 hepatocellular carcinoma cells, the proliferation ability of Huh7 cells was significantly inhibited in vitro, and the tumor growth rate of nude mice was slowed down, and the activity of JAK-STAT pathway in Huh7 cells was decreased, and the levels of c-MYC and Bcl-xl protein were decreased. In addition, deletion of CXCR3 can effectively inhibit IL-6-mediated JAK-STAT pathway activation.@*Conclusion@#CXCR3 high expression indicated that the survival rate was poor, and the target silencing of CXCR3 gene could inhibit the proliferation of hepatocellular carcinoma cells and maybe related to inhibition of JAK-STAT pathway activity. CXCR3 may be a potential target for the treatment of hepatocellular carcinoma.

The role and mechanism of SDF-1/CXCR4 signaling pathway in rat model of chronic renal allograft rejection / 中华器官移植杂志

Objective To investigate the role and mechanism of SDF-1/CXCR4 in the development of chronic rejection (CR) in rat models.Methods CR rat models were established using Fisher 344 to Lewis rats.In the blank control group (n=10),Lewis rats getting isotransplantation were treated with Cyclosporine A.CR rat models were established in positive group (n=10) and the rats were treated with Cyclosporine A.CR rat models were also established in CXCR4 antagonism group (n=10) and the rats were treated with both Cyclosporine A and AMD3100 (1 mg/kg).The serum creatinine levels were monitored every week.Kidney grafts were harvested 12 weeks after transplantation for histological analysis.We evaluated graft injuries using chronic allograft damage index (CADI) scores.Q-PCR and Western blotting were used to measure CXCR4,TGF-β1/Smad3 signaling pathway and α-smooth muscle actin (α-SMA) expression in renal allograft tissues.Results The serum creatinine levels in blank control group and CXCR4 antagonism group were significantly lower than those in positive control group (P<0.05).The blank control group and CXCR4 antagonism group presented milder pathological manifestations of CR.The CADI score in CXCR4 antagonism group was 3.54,which was lower than that of positive control group (P<0.05).The expression of biological markers in TGF-β1/Smad3 signaling pathway and SDF-1/CXCR4 signaling pathway was significantly lower in blank control group and CXCR4 antagonism group than in positive control group (P<0.05).Conclusion SDF-1/CXCR4 signaling pathway may play a crucial role in the development of CR.The usage of SDF-1/CXCR4 antagonist can protect renal allograft by inhibiting the TGF-β1/Smad3 pathway.Therefore,antagonism of CXCR4 may provide a novel way to prevent the development of CR.

Chemokine CXCL12 and its receptor CXCR4 expression and significance in hypopharynx cancer / 中国医师杂志

Objective To investigate the chemokine 12 (CXCL12) and chemokine receptor 4 (CXCR4) expressions in hypopharyngeal carcinoma and its place in the disease development,invasion and metastasis of significance.Methods Immunohistochemistry was used to detect the expressions of CXCL12 and CXCR4 in 35 cases of hypopharyngeal cancer tissues and in 28 cases of tumor-adjacent non-tumor tissues.Results The expressions of CXCL12 and CXCR4 in the hypopharynx carcinomas were significantly higher (P ＜ 0.05).Both expressed in hypopharyngeal carcinomas was significantly positively correlated (P ＜ 0.01).Both hypopharynx cancer in lymph node metastasis group were significantly higher than the expression of cervical lymph node metastasis group,the difference was significant (P ＜ 0.05).Conclusions CXCL12 and CXCR4 are involved in hypopharynx cancer development,invasion and metastasis,and there is a positive feedback regulation mechanism between two factors.Moreover,CXCL12 and CXCR4 have synergistic effect in development,invasion and metastasis of hypopharynx cancers.

Isolation and culture of corneal neovascular endothelial cells and expression of chemokine receptors / 中华实验眼科杂志

Background The pathogenesis and mechanism research of corneal neovascularization is of important significance for the prevention and management of corneal neovascularization.Some relative researches are being performed on non-corneal neovascularization-derived vascular endothelial cells, so the results are affected to a certain extent.Objective This study was to isolate and culture vascular endothelial cells from experimental corneal neovascularization tissue and detect the expression of chemokine receptors in vitro.Methods Corneal neovascularization models were established on 10 SPF male BALB/c mice with the age of 7-8 weeks by sticking the filter papers with NaOH on the central corneas, and then the immunofluorescence technique was use to assay the CD31 expression in corneal flatmount 2 weeks after modeling.Corneal pieces were made in 2 weeks after alkali burn and then were digested by collagenase type D.Vascular endothelial cells were isolated from neovascularized tissue by affinity purification using magnetic beads coated with anti-CD31.The cells were cultured on fibronectin-coated walls and then identified by immunocytochemistry.Reverse transcription-PCR was employed to detect the expressions of chemokine receptors in the cells.The use and care of the animals complied with ARVO Statement and this experimental procedure was approved by Soochow University Animal Care Committee.Results Corneal neovascularization occurred at 7 days and peaked at 2 weeks after modeling, and immunofluorescence exhibited the green network-like fluorescence for CD31 antibody in corneas.The cells grew against the wall 2 hours after culture with the polygon shape and large dimension, and the growth obviously quickened after passage.The cultured cells showed the positive response for CD31 antibody, showing the brown dye in cytoplasm,in contrast,the expression of CD31 was absent in corneal stromal cells.Chemokine receptors were positively expressed in the cells with the strongest expression levels in CCR1 ,CCR2,CCR3 and CCR4 mRNA and the weakest expression levels in CCR9,CXCR4 and CXCR5 mRNA,while CXCR3, CCR6, CCR10 and CX3CR1 mRNA were expressed with the moderate intensity.Conclusions Vascular endothelial cells can be obtained from experimental neovascularized corneas by affinity purification and express chemokine receptors,which facilitate the study of their biological properties.

Chemokines and lung cancer / 国际肿瘤学杂志

Chemokines and chemokine receptors involve in biological activity and pathological process widely.It has been reported that many tumor cells overexpress functional chemokines.In lung cancer,chemo-kines involve in its proliferation,apoptosis,invasion and metastasis.Chemokines and chemokine receptor over-expressed in lung cancer can be used as specific target for pertinent anti-tumor treatment.

Expression and clinical significance of serum chemokines in patients with lung cancer / 国际肿瘤学杂志

Objective To detect the expression levels of multiple serum chemokines including IFN-inducible T cell chemoattractant (ITAC),Fractalkine,macrophage inflammatory protein (MIP)-3α,IL-8,MIP-lα,MIP-1β in patients with lung cancer and explore their association with the clinical characteristics of lung cancer as well as the correlations among these chemokines.Methods Forty newly diagnosed patients with lung cancer and thirty healthy controls were enrolled for detection of the serum levels of 6 kinds of chemokines by Luminex technology.The correlations of clinical characteristics of lung cancer with these chemokines and the correlations among these chemokines were analyzed by SPSS 17.0 software.Results The serum levels [M (QR)] of IL-8,Fractalkine and MIP-3α in patients with lung cancer were 5.16 (4.74),128.45 (141.89),10.31 (8.88) respectively,and 2.01 (0.95),61.46 (74.81),8.08 (5.87) respectively in control group,with significant differences (Z =-4.783,P ＜0.001;Z =-4.046,P ＜0.001;Z =-3.105,P =0.002).The expression of MIP-1β in lung adenocarcinoma was significantly higher than that in squamous carcinoma [18.32 (12.27) vs.13.72 (7.31),Z =-2.212,P =0.027],and of ITAC in squamous carcinoma was significantly higher than that in small cell lung cancer [24.51 (22.48) vs.9.28 (4.85),Z =-2.460,P =0.014].The expressions of MIP-3α and Fractalkine were positively correlated in the two groups (r =0.619,P＜0.001;r=0.766,P＜0.001).Conclusion The expressions of IL-8,Fractalkine and MIP-3α increase significantly in lung cancer patients,and they are may play important roles in metastatic lung cancer.

Synthesis of 6-18F-Py-AMD3465 and the microPET/CT imaging of this agent in mice bearing A549 tumor / 中华核医学与分子影像杂志

Objective To synthesize 628F-Py-AMD3465,to investigate its biodistribution in mice and to perform the microPET/CT imaging on mice bearing human lung cancer cell (A549).Methods AMD3465 quaternary ammonium salt precursor was directly labeled with 18F,then 628F-Py-AMD3465 was synthesized through nucleophilic reaction,hydrolysis,neutralization and the product was purified using HPLC.The labeling yield and radiochemical purity were analyzed by HPLC.Fifteen Kunming mice were injected with 5.55 MBq of 628F-Py-AMD3465 and sacrificed at 5,20,40,60 and 120 min postinjection.The selected tissues were harvested and weighed,and the radioactivity in the tissues was measured by an automated γ-spectrometer.The ％ID/g was calculated.MicroPET/CT studies were performed on A549-bearing mice after injecting 6-18F-Py-AMD3465 through vena caudal.Paired t test was used.Results 6-18F-Py-AMD3465 was successfully synthesized with the labeling yield of (9.0±2.0)％,the total synthesis time was about 60 min,and the radiochemical purity was more than 98％.Biodistribution studies showed that the radiouptake was higher in the kidneys and bladder of normal mice,which demonstrated that 6-18 F-Py-AMD3465 was mainly excreted through the kidneys.Biodistribution in A549-bearing mice was similar to that in normal mice.The tumor/muscle ratio at 40 min was 5.0,but the radiouptake of the tumor was still lower than that of the normal lung:(8.05±0.35) ％ID/g vs (9.33±0.66) ％ID/g;t=5.26,P＜0.05.MicroPET/CT imaging showed that the high-uptake location of 6-18F-Py-AMD3465 in tumor-bearing mice was similar to the normal mice,and the tumor uptake reached the maximum level at 45 min post-injection (SUV 0.67).Conclusions 6-18F-Py-AMD3465 can be synthesized by a simple method.A lower uptake could be shown in the tumor compared to that in the lung and the tracer has limited diagnostic value for lung cancer.

Induction of CCR7 gene modified immature dendritic cells to immune tolerance in rats high-risk corneal allograft / 中华实验眼科杂志

Background Graft rejection is a primary cause of corneal transplantation failure,especially in high-risk keratoplasty.How to extend the survival time of graft is a problem to be solved.Objective This study was to investigate the influence of immune tolerance on high-risk rat keratoplasty induced by donor bone marrowderived immature dendritic cells (imDCs) transfected by chemokine receptor 7 (CCR7) recombinant adenovirus (Ad).Methods Bone marrow-derived imDCs were isolated and cultured from femur marrow of one male Wistar donor rat.The cells were transfected using recombinant Ad vector with rat CCR7 gene and resuspended in 500 μl PBS containing 1％ fetal bovine serum with the cells 1 × 107.High-risk corneal transplantaion models were established using monolateral corneal alkali-burn method in 60 SD rat recipients, and then allograft keratoplasty was performed with the 30 Wistar rats as donors.The models were randomized into the PBS group,imDCs group,imDCs with blank Ad vector group and imDCs with Ad-CCR7 group following the corresponding solution injection via caudal vein on preoperative day 7 and postoperative day 3 respectively.The survival time of graft was evaluated under the slit lamp microscope once per day.On the 14th day after operation, corneal sections were prepared from 6 eyes of each group for the pathological examination,and the relative expression levels of T helper cell 1 (Th1)-related factors,interleukin-2 (IL-2) mRNA and interferon-γ(IFN-γ) mRNA,as well as Th2-related factors, IL-4 mRNA and IL-10 mRNA, were detected by reverse transcription PCR (RT-PCR).The use and care of animals complied with the ARVO statement Results The mean survival time of grafts was (10.44±1.88) , (16.00±2.18) , (15.11±2.03) and(23.67±2.83) days in the PBS group,imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group, respectively, with a significant difference among the 4 groups (F =53.005, P =0.000).Compared with the PBS group, the mean survival time of grafts was considerably extended in the imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group (t=5.220,4.385,12.423 ,all at P=0.000) ,and a remarkble prolongation of graft survival duration was seen in the imDCs with Ad-CCR7 group in comparison with the imDCs group and the imDCs with blank Ad group (t =7.204,8.039,both at P=0.000).The relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were significantly lower,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were significantly higher in the imDCs group,imDCs with blank Ad group and the imDCs with Ad-CCR7 group than those in the PBS group (all at P＜0.05).Compared with the imDCs group and the imDCs with blank Ad group, the relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were remarkably delined,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were remarkably elevated in the imDCs with Ad-CCR7 group (all at P =0.000).Conclusions Application of imDCs transfected with CCR7 recombinant Ad via vena caudalis can prolong the survival time of grafts after keratoplasty of SD rats probably by affecting the balance of Th1/Th2 cytokines.