Objective To investigate lymph node metastasis on the middle and lower segment of lⅡ and Ⅲ stage of esophageal squamous cell carcinomas,understand the factors influencing the lymph node metastasis,and provide the basis for the key areas of lymph node cleaning.Methods A retrospective study was made on the specimens of 186 patients who were middle and lower segment of Ⅱ and Ⅲ stage of esophageal squamous cell carcinomas,who had underwent radical operation through left thoracic,thoraco abdominal two field lymph node cleaning.All the cases were patients from April 2010 to December 2013 at the Inner Mongolia Medical University Clinical Medical College of Chifeng.Results A percentage (67.9％) of patients (126/186) was found with lymph node metastasis.A total of 4259 lymph node was dissected,with an averaged cleaning of (22.9 ± 8.1) lymph nodes for each case.A total of 622 lymph nodes (14.6％ =622/4 259) existed metastasis.The rate of mediastinum metastasis for middle and lower segment of esophageal cancer was 56.1％ and 16.5％,respectively.The rate of metastasis to the lower mediastinal lymph nodes was 34.6％ and 54.4％,respectively.The rate of metastasis to the celiac lymph nodes was 23.4％ and 46.8％,respectively.A significant difference was found in the metastasis locations of middle and lower segment of esophageal carcinomas (P ＜ 0.05).The top three locations of lymph node metastasis in the middle segment of esophageal squamous cell carcinomas were the lymph nodes of left artery paraesophageal,carina,and gastric bypass.The top three locations of lymph node metastasis in the lower segment of esophageal squamous cell carcinomas were paraesophageal,cardia,and gastric lymph nodes.The depths of tumor invasion,differentiation,intravascular cancer embolus were statistically significant effect on lymph node metastasis rate (P ＜ 0.05).Tumor location,and lesion length had no significant effect on the lymph node metastasis rate (P ＞ 0.05).Conclusions The lower segment of Ⅱ,Ⅲ stage esophageal squamous cell carcinoma with lymph node metastasis occurs in the lower mediastinal and abdominal lymph nodes.The middle segment Ⅱ,Ⅲ stage esophageal squamous cell carcinoma with lymph node metastasis occurs in the thoracic and abdominal lymph nodes with Jump transfer characteristics.The lymph node cleaning of the mid-dle segment includes the left artery near the stomach,paraesophageal,and carina lymph node.The lymph node cleaning of the lower segment includes paraesophageal,cardia,and gastric lymph nodes.The metastasis rate of vascular tumor thrombus is related to the depth of tumor invasion and differentiation degree.

Objective: To evaluate the antimicrobial activity of nonequilibrium plasma against Enterococcus faecalis (Ef) biofilms in vitro and to obtain novel evidence of root canal disinfection with nonequilibrium plasma. Methods: Sterile cover slips and single-rooted canals were filled with Ef and incubated to form 1-week-old and 3-week-old biofilms, respectively. The infected samples were subjected to nonequilibrium plasma, 2% chlorhexidine digluconate (CHX) and saline for 3, 10 and 30 minutes, respectively. After treatment, the killing effectiveness of nonequilibrium plasma was analyzed by using laser scanning confocal microscopy (LSCM) and colony forming unit (CFU) counting. Results: The 3-dimentional reconstruction LSCM images showed that about 48.3%-79.8% of 1-week-old Ef biofilm cells and 40.0%-67.4% of 3-week-old biofilm cells were killed by nonequilibrium plasma and 2% CHX compared to saline (P<0.05). The proportion of killing activity was lower after 3 minutes (40.0%-50.9% killing) than after 10 minutes (65.3%-77.8% killing) and 30 minutes (66.4%-79.8% killing) (P<0.05). And the killing of biofilm bacteria was fastest during the first 3 minutes (13.3%-17.0% killing per minute) and slow down greatly after 10 minutes. Remarkably more bacteria were killed in 1-week-old Ef biofilms (48.3%-79.8% killing) than in 3-week-old biofilms (P<0.05). Conclusions The nonequilibrium plasma killed more Ef biofilm cells in infected root canals showed promotional as an additional approach against bacterial biofilms during root canal disinfection.

Objective: To evaluate the antiseptic effect of combined using of 5% sodium hypochlorite and calcium silicate-based root canal sealer against Enterococcus faecalis (Ef) biofilms in infected dentinal tubules in vitro. Methods: Cells of Ef were inoculated into the dentinal tubules of single-rooted teeth (without caries, periapical lesions and malformations extracted due to periodontal disease or orthodontic reasons; collected from Department of Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University) with centrifugation and incubated in brain-heart infusion (BHI) to form 3-week-old biofilms. The infected samples were subjected to sodium hypochlorite or sterile water bathing for 10 minutes followed by calcium silicate-based root canal sealer (iRoot SP) (calcium silicate-based group), Gutta-percha group and sterile water group placed on the root canal wall for 1, 4 and 12 weeks. There were two samples in each treatment at each point. The antiseptic effectiveness of combined use of sodium hypochlorite and calcium silicate-based root canal sealer was analyzed by laser scanning confocal microscope (LSCM), ANOVA and LSD-t test. Results: After treatment with 5% sodium hypochlorite, in calcium silicate-based group for 4 and 12 weeks more Ef biofilm cells [(75.3±3.5)% and (74.8±3.8)%] were killed than in Gutta-percha group [(65.9±4.1)% and (63.0±3.7)%] and sterile water group [(63.9±4.0)% and (64.2±3.5)%] (P<0.05). After being treated with sterile water, the proportion of dead bacterial cells in calcium silicate-based group for 1, 4 and 12 weeks [(27.5±4.6)%, (43.0±4.4)% and (40.3±6.1)%] were more than those in Gutta-percha group and sterile water group (P<0.05). After being treated with 5% sodium hypochlorite or sterile water, more biofilm bacteria were killed in calcium silicate-based group for 4 and 12 weeks than in calcium silicate-based group for 1 week (P<0.05). Conclusions The combined use of sodium hypochlorite and calcium silicate-based root canal sealer kills more biofilm cells in infected dentinal tubules.

OBJECTIVETo judge whether algesia sensitization of some acupoints is existed and whether the acupoint algesia sensitization area is expanded in the patients of intestinal cancer.

METHODSTotally, 30 patients of intestinal cancer and 30 healthy subjects were included. The electronic Von Fray was used to determine the pressure-pain thresholds at 13 acupoints relevant with gastrointestinal disorders and the reference points at the sites 1and 2lateral to those points as well as the sites at the corresponding nerve segments. Compared with the pressure-pain thresholds at the reference points of the different segments, the relative value was calculated. The changes were analyzed in the pressure-pain thresholds at the relevant acupoints on the body surface in the patients of intestinal cancer as compared with the relative pressure-pain thresholds in the healthy volunteers.

RESULTSThe pressure-pain thresholds at Zusanli (ST 36), Shangjuxu (ST 37), Xiajuxu (ST 39), Quchi (LI 11) and Dachangshu (BL 25) in the patients of intestinal cancer were all significantly reduced as compared with those of the healthy subjects (<0.05,<0.01,<0.001). At the non-acupoint sites 1and 2lateral to those acupoints as well as at the sites of the same segments, the pressure-pain thresholds were reduced significantly as compared with the control group (<0.05,<0.01,<0.001). Particularly, the sensitization zone of Yinlingquan (SP 9) focused on the acupoint, the site 1lateral to it as well as the non-acupoint sites of the same segments (<0.01,<0.001).

CONCLUSIONThe acupoint sensitization is displayed at Zusanli (ST 36), Shangjuxu (ST 37), Xiajuxu (ST 39), Quchi (LI 11), Dachangshu (BL 25) and Yinlingquan (SP 9) and the sensitization area is expended in the patients of intestinal cancer.

Objective: To establish an inducible macrophage-specific knockout G protein-coupled receptor kinase 2 ( GRK2) gene ( GRK2flox / flox Lyz2-CreERT + ) mice model． Methods: GRK2flox / flox Lyz2-CreERT + mice were constructed based on Cre / LoxP system．The genotypes of GRK2flox / floxLyz2-CreERT + mice were identified by PCR amplification and agarose gel electrophoresis．After the mice were sacrificed by carbon dioxide method，the expression of GRK2 in bone marrow-derived macrophages ( BMDMs) and peritoneal macrophages ( PMs) was detected by Western blot．Immunofluorescence was used to detect GRK2 expression in mouse brain，heart and spleen macrophages．The M1 / M2 ratio in PMs induced by prostaglandin E2 (PGE2) was analyzed by flow cytometry. Results: The results of genotype identification showed that the mice with a band at 355 bp in the length of the flox amplification product and a band at 355 bp in the length of the Cre amplification product were GRK2flox / floxLyz2-CreERT + mice．Western blot results showed that GRK2 expression in BMDMs and PMs of GRK2flox / floxLyz2-CreERT + mice decreased compared with GRK2flox / flox mice(P＜0. 01) ．Immunofluorescence results showed that GRK2 expression decreased in the brain，heart and spleen of GRK2flox / floxLyz2-CreERT + mice compared with GRK2flox / flox mice (P < 0. 01) ．Flow cytometry showed that compared with GRK2flox / flox mice，there was no significant difference in the proportion of CD86 / CD206 in the PMs of GRK2flox / floxLyz2-CreERT + mice．Under PGE2 ( 10 μmol / L) stimulation， the proportion of CD86 / CD206 in GRK2flox / floxLyz2-CreERT + mice PMs increased (P ＜0. 01) ．The proportion of CD86 / CD206 in the PMs of GRK2flox / flox mice was higher than that of GRK2flox / floxLyz2-CreERT + mice(P＜0. 01) . Conclusion In this study，GRK2flox / floxLyz2-CreERT + mice model was successfully constructed，and the mice promoted PGE2-induced polarization of PMs to M2-type macrophages compared with control mice.

Rheumatoid arthritis (RA) is an autoimmune disease with a complex etiology. Monocyte-derived macrophages (MDMs) infiltration are associated with RA severity. We have reported the deletion of G-protein-coupled receptor kinase 2 (GRK2) reprograms macrophages toward an anti-inflammatory phenotype by recovering G-protein-coupled receptor signaling. However, as more GRK2-interacting proteins were discovered, the GRK2 interactome mechanisms in RA have been understudied. Thus, in the collagen-induced arthritis mouse model, we performed genetic GRK2 deletion using GRK2^f/fLyz2-Cre^+/- mice. Synovial inflammation and M1 polarization were improved in GRK2^f/fLyz2-Cre^+/- mice. Supporting experiments with RNA-seq and dual-luciferase reporter assays identified peroxisome proliferator-activated receptor γ (PPARγ) as a new GRK2-interacting protein. We further confirmed that fms-related tyrosine kinase 1 (Flt-1), which promoted macrophage migration to induce angiogenesis, was inhibited by GRK2-PPARγ signaling. Mechanistically, excess GRK2 membrane recruitment in CIA MDMs reduced the activation of PPARγ ligand-binding domain and enhanced Flt-1 transcription. Furthermore, the treatment of mice with GRK2 activity inhibitor resulted in significantly diminished CIA pathology, Flt-1⁺ macrophages induced-synovial inflammation, and angiogenesis. Altogether, we anticipate to facilitate the elucidation of previously unappreciated details of GRK2-specific intracellular signaling. Targeting GRK2 activity is a viable strategy to inhibit MDMs infiltration, affording a distinct way to control joint inflammation and angiogenesis of RA.