Objective To investigate the effects of active vitamin D (VD) on the expression of triggering receptor expressed on myeloid cells-1 (TREM-1) in renal tissue of diabetic nephropathies (DN) rats and to explore the impact of TREM-1 on adhesion and migration capacity of macrophage.Methods DN rat models were established by streptozotocin.Rats were randomly distributed into four groups:control (NC) group,VD group,DN group and DN+VD group (DN rats with 0.1 μg · kg-1 · d-1 calcitriol by garages).Rats were sacrificed respectively at 8 weeks and 12 weeks after treatment.Pathological changes in kidney tissue were detected and the expressions of CD68 and TREM-1 were acquired by immunohistochemistry stain and Western blotting.In vitro,RAW264.7 cells were divided into NC group,VD group,high glucose (HG) group and HG+VD group.In HG+VD group rats were treated by high glucose with 10-8 mol/L 1,25(OH)2D3.TREM-1 expression was measured by immunohistochemistry stain and Western blotting,and the ability of macrophage in migration and adhesion was evaluated by Transwell migration assay and adhesion assay.TREM-1 siRNA was transferred to silence TREM-1 expression,while plasmid of TREM-1 was transferred for high expression.Their ability of adhesion and migration in macrophage and the effect of 1,25(OH)2D3 were examined.Results (1) Compared with the NC group,the expressions of CD68 and TREM-1 were increased in DN group (P ＜ 0.05),whereas markedly decreased in DN+VD group (P ＜ 0.05).(2) The number of adhesion and migration cells,and the expression of TREM-1 protein in macrophage were obviously increased in HG group as compared with those in NC group (all P ＜ 0.05);whereas above changes were markedly decreased in HG+VD group than those in HG group (P ＜ 0.05).(3) The number of adhesion and migrated macrophage was reduced after TREM-1 siRNA intervention (all P ＜ 0.05).VD could significantly decrease the effect of high glucose on adhesion and migrated macrophages after TREM-1 siRNA (all P ＜ 0.05).(4) Adhesion and migration of macrophage were increased via TREM-1 overexpression (all P ＜ 0.05),but the effects of VD on high glucose-induced adhesion and migration of macrophage were disappeared.Conclusions VD can suppress the adhesion and migration of macrophage via reducing the expression of TREM-1,and inhibit infiltration of macrophage in renal tissue of DN rats.

Objective To investigate the effect of macrophages on podocytes apoptosis in diabetic nephropathy. Methods Differentiated mouse macrophages (RAW264.7) were exposed to normal glucose, high glucose, then the conditioned media (CM) was collected and considered as NC-CM or HG-CM, respectively. Western blotting and immunofluorescent staining were used to detect the specific markers for M1 macrophages (iNOS) and M2 macrophages (MR). ELISA was used to detect the concentration of TNF-α in the CM. Then normal PRMI 1640 media (control), NC-CM or HG-CM was added to podocytes. In some experiments, ROS inhibitors (Tempo), p38 MAPK inhibitor (SB203580), anti-TNF-α neutralizing antibody, and IgG1 isotype control were respectively added to cells with HG-CM. Besides, recombinant mouse TNF-α alone was applied to incubate podocytes. Podocytes apoptosis was accessed by Annexin V-FITC/PI and Hoechst33342 staining. DCFH-DA staining was used to analyse ROS level. Western blotting was used to detect cleaved casepase-3, p38MAPK, and p-p38MAPK protein. Results Macrophages were activated when exposed to high glucose, displaying pro-inflammatory M1 polarization with higher iNOS and lower MR expression. HG-CM but not NC-CM trigged podocytes apoptosis, up-regulated ROS, cleaved casepase-3 and p-p38MAPK. However, the podocytes apoptosis trigged by HG-CM was abolished by either a ROS inhibitor (Tempo) or a p38 MAPK inhibitor (SB203580). Additionally, TNF-α was increased in the HG-CM. TNF-α protein in macrophage was aslo increased when exposed to high glucose. Anti-TNF-α neutralizing antibody blunted the apoptotic response, excess ROS generation and p-p38 MPAK expression in podocytes induced by HG-CM. Moreover, addition of recombinant TNF-α similarly led to podocytes apoptosis, increased ROS and p38 MPAK expression. Conclusion M1 macrophages activated by high glucose released TNF-α to promote podocytes apoptosis via ROS-p38 MAPK pathway.

Objective To establish a canine model of intervertebral disc extrusion by surgery and observe the histological and microcirculatory changes of the spinal cord , in order to accumulate data for studies on the pathology and mechanism of treatment for intervertebral disc extrusion .Methods Normal healthy adult dogs were divided randomly into two groups:normal control group and model group .To simulate the intervertebral disc extrusion caused by spinal cord compression, 6Fr double lumen catheter with ballon was inserted into the spinal cord T 12-T13 and filled with about 5 mL Iohexol after the exposure of spinal cord L 1 by hemilaminectomy .The spinal cord blood flow ( SCBF) at the L1 level before and after compression was measured by laser-Doppler flowmetry .Morphological changes of the compressed spinal cord at 14 days after compression was examined by histopathology .Results The ( Texas spinal cord injury score ) ( TSCIS) scores of the motor function of bilateral hind limbs were highly significantly decreased (P<0.01).The blood flow of spinal cord at the L1 level was significantly decreased (P<0.05) after compression than that before .Compared with the normal control group, the model group showed abnormal vacuolization in the white matter and the number of normal neurons in the ventral horn of gray matter was significantly lower ( P<0.01 ) .Conclusions Our findings demonstrate that canine models of intervertebral disc extrusion can be successfully established by balloon catheter compression , showing local impairment of microcirculation and histological changes in the spinal cord .This canine model may provide a useful model for evaluation of therapeutic effects of acupuncture and for mechanism studies .

The potential antiviral and anti-inflammatory mechanism of Anti-601 Mixture, a traditional Chinese medicine compound preparation, was studied by network pharmacology and molecular docking. The chemical constituents and targets of astragali radix, phellodendri chinensis cortex, rhei radix et rhizome, isatidis radix and lonicerae japonicae flos in Anti-601 Mixture were searched by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). The genes corresponding to the target were searched through the UniProt database, and the drug-compound-target (gene) network was constructed by Cytoscape 3.7.2. Then the functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were conducted through Webgestalt to predict the mechanism. The network of drug-compound-target (gene) contained 5 drugs, 100 compounds and 207 targets. Functional enrichment analysis resulted in 717 GO items (P≤0.05), among which 240 were biological process (BP) items, 240 were cell composition (CC) items, and 237 were molecular function (MF) items. The 209 signaling pathways were obtained by enrichment screening of KEGG pathway (P≤0.05). Molecular docking showed that the active ingredients of Anti-601 Mixture, such as Stigmasterol, quercetin, luteolin, acacetin, β-sitosterol, kaempferol,had strong affinity with PTGS2 target. Active compounds in Anti-601 Mixture may regulate multiple signaling pathways including advanced glycation end products and its receptor (AGE-RAGE), IL-17, tumor necrosis factor (TNF) through target of prostaglandin-endoperoxidase synthase 2(PTGS2), thus playing an antiviral and anti-inflammatory role.

Objective To observe the impact factors of the diagnostic accuracy of fine needle aspiration biopsy(FNAB)for papillary thyroid carcinoma(PTC).Methods Totally 468 patients with single PTC confirmed by postoperative pathology who underwent FNAB before surgery were enrolled.The impact of clinica,l ultrasonic and pathological features on the accuracy of FNAB diagnosis were analyzed.Results The accuracy of FNAB for diagnosing PTC was 71.37%(334/468).The maximum diameter and location of PTC were both impact factors of the diagnostic accuracy of FNAB.The maximum diameter of 0.7 cm was the optimal cutoff value of FNAB for diagnosing PTC,and the diagnostic accuracy of FNAB for PTC with the maximum diameter＜0.7 cm and those≥0.7 cm was 62.96%(119/189)and 77.06%(215/279),respectively.The diagnostic accuracy of FNAB for PTC located in the difficult and easy area of puncture was 52.53%(52/99)and 76.42%(282/369),respectively.The diagnostic accuracy of FNAB for PTC with the maximum diameter≥0.7 cm and located in the easy area,≥0.7 cm and located in the difficult area,＜0.7 cm and located in the easy area,＜0.7 cm and located in the difficult area was 80.43%(185/230),61.22%(30/49),69.78%(97/139)and 44.00%(22/50),respectively.Conclusion The maximum diameter and location of PTC were both impact factors of the diagnostic accuracy of FNAB.