1.Production and mechanism of CCL5 by macrophages in U14 cervical cancer-bearing mice during infection
Hong REN ; Guoli REN ; Limin SUN ; Xiuhua FAN ; Yuran WANG ; Xiaoxi LI
Chinese Journal of Obstetrics and Gynecology 2015;(5):367-373
Objective To investigate the production and mechanism of chemokine (C-C motif) ligand 5 (CCL5) by macrophages in U14 cervical cancer-bearing mice during infection. Methods The U14 cervical cancer cells were injected in C57BL/6 mice to induce tumor-bearing condition. Lipopolysaccharide (LPS) was injected into C57BL/6 mice to induce infection. The protein expression of CCL5 in the serum and the CCL5 mRNA expression in inflammatory cells were measured by ELISA and fluorescence quantitative-PCR in four groups. Macrophages were induced in the tumor conditioned medium (TCM) which extracted from mice serum. The protein expression levels of CCL5, prostaglandin E2 (PGE2) and cyclic adenosine monophosphate (cAMP) in the medium and CCL5, PGE2 and cAMP mRNA expression in the macrophages were detected in different groups. In order to determine whether the inhibition was related to PGE2, selective cyclooxygenase 2(COX-2) inhibitor NS398 was used to reverse this phenomenon and protein kinase A (PKA) inhibitor H89 demonstrated the mechanism through blocking cAMP/PKA signaling pathway. Results (1) The protein and mRNA level of CCL5 in tumor-bearing mice were respectively (151±35) pg/ml and 1.0, which were lower than those in the tumor-free mice (691 ± 85) pg/ml and 4.5 ± 0.8, there were significant difference between them (all P<0.05). The protein and mRNA level of PGE2 in tumor-bearing mice were (1 198±83) pg/ml and 5.8±0.8, which were higher than those in the tumor-free mice (187±25) pg/ml and 1.0, the difference were significant (all P<0.05). The protein and mRNA level of CCL5 in tumor-free+LPS mice were (4 049±141) pg/ml and 31.5±2.0, which were higher than those in the tumor-bearing+LPS mice (1 951±71) pg/ml and 12.1±2.8, the difference were also significant (P<0.05). The protein and mRNA level of PGE2 in tumor-free+LPS mice were (676±70) pg/ml and 3.4±0.4, which were lower than those in tumor-bearing+LPS mice (2 550±382) pg/ml and 11.6±0.9, the difference were also significant (all P<0.05). (2) Macrophages were cultured in vitro using TCM derived from mice. The protein and mRNA level of CCL5 in tumor-bearing mice TCM were respectively (1 626 ± 177) pg/ml and 28.6 ± 1.2, which were higher than those in the tumor-free mice TCM [(27 ± 3) pg/ml and 1.0], there were significant difference (P<0.05). The protein and mRNA level of PGE2 in tumor-bearing mice TCM were (790 ± 156) pg/ml and 1.7 ± 0.3, which were higher than those in the tumor-free mice TCM [(448 ± 115) pg/ml, 1.0], the difference were significant (all P<0.05). The protein and mRNA level of cAMP in tumor-bearing mice TCM were (164 ± 30) pg/ml and 1.6 ± 0.3, which weres higher than those in the tumor-free mice TCM [(118 ± 25) pg/ml,1.0], the difference were significant (all P<0.05). The protein and mRNA level of CCL5 in tumor-free + LPS mice TCM were (10 475 ± 742) pg/ml and 212.0 ± 5.7, which were higher than those in the tumor-bearing+LPS mice TCM [(6 375±530) pg/ml, 142.3±2.5], the difference were significant (all P<0.05). The protein and mRNA level of PGE2 in tumor-free+LPS mice TCM were (2 438±95) pg/ml and 4.3±0.7, which weres lower than those in the tumor-bearing + LPS mice TCM [(3 441 ± 163) pg/ml, 5.9 ± 0.3], the difference were significant (all P<0.05). The protein and mRNA level of cAMP in tumor-free+LPS mice TCM were (340 ± 13) pg/ml and 4.1 ± 0.4, which were lower than those in the tumor-bearing + LPS mice TCM [(542 ± 42) pg/ml, 5.4 ± 0.5], the difference were significant (all P<0.05). (3) Using COX-2 inhibitor NS398 in the tumor-bearing+LPS mice, the protein and mRNA level of CCL5, PGE2 and cAMP were (7 691±269) pg/ml and 159.0±8.9, (2 820±152) pg/ml and 4.9 ± 0.3, (465 ± 8) pg/ml and 4.3 ± 0.4, respectively, and there were significant difference (all P<0.05), compared to before treatment. Using PKA inhibitor H89 in the tumor-bearing+LPS mice, the protein and mRNA level of CCL5, PGE2 and cAMP were (8 375±520) pg/ml and 177.0±8.8, (2 650±35) pg/ml and 4.7 ± 0.4, (368 ± 13) pg/ml and 3.1 ± 0.7, respectively, and there were significant difference (all P<0.05), compared to before treatment. Conclusion TCM of U14 cells activated macrophages to release PGE2 could inhibit the expression of CCL5 levels by cAMP/PKA signaling pathway.
2.3.0T MR Susceptibility-weighted Imaging for Substantia Nigra Pars Compacta in Primary Parkinson’s Disease
Bo WANG ; Hongfei AN ; Jie ZHANG ; Guoli BI ; Kunhua WU ; Lixiang REN ; Ling SHEN
Journal of Kunming Medical University 2014;(2):27-30
Objective To evaluate the application of measurement of T2*value,width of substantia nigra pars compacta (SNc) and the ratio of the width to the midbrain diameter in diagnosing Parkinson disease (PD) in early stage with susceptibility weighted imaging ( SWI) by 3T MR. Methods 59 patients with early stage idiopathic PD patients and 59 healthy controls,ranging in same ages and gender,had been scanned with routine sequences and SWI sequences by 3T MR. T2*value,width and the ratio of the width to the midbrain diameter of SNc were measured. The results of measurement were analyzed and compared. Results (1) The T2*values, width and the ratio of the width to the midbrain diameter was decreased in homolateral side SNc of symptoms of subjects with PD compared with the healthy controls ( <0.05) . (2) There was a significant reduction in the T2*values and the ratio of the width to the midbrain diameter in contralateral side SNc symptoms of subjects with PD compared with the healthy controls (P<0.05) . There was no differences in width of SNc ( >0.05) . Conclusion Measurement of T2*value, width and the ratio of the width to the midbrain diameter of SNc with SWI is reliable to diagnose PD.
3.MRI diagnosis of single lesion in the corpus callosum department
Guoli BI ; Xiarong GONG ; Kunhua WU ; Ying ZHAO ; Maoping RUI ; Lixiang REN ; Yuhui CHEN
Journal of Practical Radiology 2016;(2):185-187
Objective To evaluate MRI diagnostic value for single lesion characteristics in the splenium of corpus callosum.Methods MRI features,clinical data,and parts of follow-up results of 9 cases with single lesion in the splenium of corpus callosum were analyzed retrospectively.Results (1)Clinical manifestations:headache and dizziness occurred in 4 cases,syncope in 3 cases,fever in 2 cases, physical activity barriers in 2 cases.(2)Clinical diagnosis:hypoglycemic encephalopathy were rescaned one month later in 3 cases, in which the previous lesion completely disappeared.Clinical experience of encephalitis were improved after treatment in 2 cases. Cerebral infarction,epilepsy,brain injury and degeneration were diagnosed respectively in each one case,in which lesion still existed after treatmented.(3)Image findings:despite the different clinical manifestations,image features of all cases were quite similar. Round or foliated like lesions of slightly long T1 and long T2 signals in the splenium of corpus callosum were presented in all cases. High signals on diffusion weighted imaging and low signals on the ADC were showed with same lesions,andno obvious enhancement after contract media injected was seen.Conclusion Single lesions in the splenium of corpus callosum are showed in many diseases. The image features of hypoglycemic encephalopathy or encephalitis have certain characteristics (single lesion is reversible).Accurate diagnosis need to combine with clinical data and medical history.