Objective To investigate effects of exogenous substance P (SP) on proliferation of humau skin fibroblast (HSF) and expression of its monocyte chemoattractant protein-1 (MCP-1) under high glucose condition.Methods HSF cultured in high glucose DMEM medium were treated with SP of various concentrations at different time points.Levels of MCP-1 in the supernatants were determined by ELISA and time-effect relationship of SP regulating expression of MCP-1 was analyzed.Expression of MCP-1 mRNA in HSF was detected by semi-quantitative RT-PCR and Real Time-PCR,Conclusion,while cxpression of MCP-1 by Western blot.Proliferation of HSF treated with SP of different concentrations was detected by cell counting Kit-8 (CCK-8).Results MCP-1 had a strong expression at 8 hours after SP disposal (P ＜0.05) and reached the peak at 24 hours (P ＜ 0.01).Meanwhile,MCP-1 expressed the most at 10 nmol/L and 100 nmol/L SP (P ＜0.01).SP significantly enhanced the expression of MCP-1 mRNA and its synthesis under high glucose condition,especially at the concentration of 10 nmol/L (P ＜0.01).Also,SP induced obvious proliferation of HSF at concentrations of 10 nmol/L and 1 000 nmol/L(P ＜ 0.05).Conclusion SP promotes proliferation of HSF and expression of MCP-1 in high glucose DMEM medium,which may be of significance in promoting diabetic wound healing.

Objective To investigate the necessity of primary neurorrhaphy (direct end-to-end anastomosis) when the recurrent laryngeal nerve(RLN) is severed during thyroid surgery.Methods 15 patients who suffered from iaotmgenic unilateral complete RLN injury or whose unilateral RLN had to be sacrificed because of disease invasion had a primary repair of RLN by direct end-to-end anastomosis.In control group,26 patients who did not have a nerve repair were enrolled into this study.Subjective evaluation of aspiration and voice quality were based on patient reports and hearer reports for all patients.9 patients with neurorrhaphy and 12 patients without nerve repair were followed with videolaryngoscopic examination.Results 14 patients undergoing neurorrhaphy restored normal voice at 2-5 months postoperatively.Although there were no significant functional motion of the vocal fold,slight adductory movement of the affected arytenoid was found with good tension vocal cords and symmetric arytenoids of the glottis during phonation.Only 2 patients without nerve repair had nearly restored normal voice.The patients with hoarseness had stiff arytenoids and atrophic folds resulting in glottal gap.Conclusions Neurorrhaphy is a simple and effective method to restore the normal aspiration and voice quality of patients with unilateral complete recurrent laryngeal nerve injuries.

Objective To study the effects of granulocyte-macrophage colony stimulating factor (GM-CSF) on the expression of vascular endotllelial growth factor (VEGF) in human dermal fibroblast. Methods In vitro human dermal fibroblasts in good status were incubated with GM-CSF (GM-CSF group) or non-GM-CSF (control group) culture medium for different periods of time. The mRNA, protein expression of VEGF in derma fibroblast were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively, and the secretion of VEGF in supernatant was measured by enzyme linked immunosorbant assay (ELISA). Results The expression of VEGF mRNA from dermal fibroblasts was increased significantly after l or more hours of incubation with GM-CSF comparing with the control (P＜0.05). 6 hours of stimulation by GM-CSF caused maximal expression of VEGF mRNA. The expression of VEGF protein in dermal fibroblasts was increased from 12 hours and was peaked at 24 hour after stimulation by GM-CSF. VEGF protein from the supernatant of the dermal fibroblasts was also raised persistently from 12 hour after stimulation by GM-CSF and was improved remarkably compared with the control. Conclusions GM-CSF can up-regulate directly the expression of VEGF in human derma fibroblast, which may be one of the mechanisms that GM-CSF accelerates neovascularization in wound healing.

Objective By proteomic analysis of differentially expressed protein profiling in pancreatic cancer using antibody microarray, new tumor marker of pancreatic cancer was supposed to be discovered. Methods The antibody microarray containing 378 monoclonal antibodies was applied to detect the differentially expressed proteins of 7 sets of pancreatic ductal adenocareinoma pooled samples and their paired normal pancreas tissues pooled samples. Results 11 up-regulated proteins (UbcH6, GABA b R2, Plakophilin 2a, Inhibitor 2, Nestin, ShcC, PRK2, Neurogenin 3, STAT 3, NHE-3 and SRP54) and 9 down- regulated proteins (DCC, HPV-16 L1, RACKI, Gelsolin, Rabaptin-5, DBP2, IKKa/I, c-Cbl, FXR2) were found in pancreatic cancer. Conclusions Antibody microarray was an effective comparative proteomic technology. These dys-regulated proteins facilitated to elucidate the mechanisms of pancreatic cancer and to identify new diagnostic markers and therapeutic targets.

Objective To investigate the expressions of the annexin A1 ( ANXA1 ) in pancreatic cancer and the correlations with proliferating cell nuclear antigen (PCNA). Methods Tissue microarray instrument was used to construct the chip. There were 39 samples of normal pancreas tissue, 42 samples of pancreatic cancer, 7 samples of islet cell tumor and 8 samples of ampullary carcinoma. The expressions of ANXA1 and PCNA protein were examined by immunohistochemistry and their correlation was analyzed. Results The off-chip rate of tissue chips after immunohistochemistry was 11.7% ～ 13.3%, which could satisfy the needs for experiment. Immunohistochemical results of pancreatic cancer tissue microarray showed that the positive rate of ANXA1 expression in pancreatic cancer was 71.4% (30/42), which was significantly higher when compared with that of normal pancreas tissue ( 18.4%, 7/38; P < 0. 01 ). The positive rate of PCNA expression in pancreatic cancer was 73.8% (31/42), which was significantly higher than that of normal pancreas tissue (22.2%, 8/36; P < 0.01 ). The expression of ANXA1 had a significant correlation with PCNA in pancreatic cancer ( P < 0.01 ). Conclusions The expression of ANXA1 protein was significantly increased in pancreatic cancer. The expression of ANXA1 was significantly correlated with the expression of PCNA.

Objective To explore the role of ubiquitin-proteasome pathway for the gelsolin protein degradation in pancreatic cancer.Methods Pancreatic cancer cell lines BxPC-3 and PANC-1 were first treated with specific 26s proteasome inhibitor lactacystin.Immunoblots of cell lysates were probed for gelsolin expression.To determine whether gelsolin was conjugated to ubiquitin,proteins extracted from the cells with or without lactacystin were immunoprecipitated with anti-gelsolin antibody,followed by Western blot analysis.Results The expression of gelsolin protein increased obviously after treatment with lactacystin in BxPC-3 cells for 12 h.Using anti-gelsolin antibody to immunoprecipitate gelsolin protein and followed by Western blot using anti-ubiquitin monoclonal antibody,it was found that inhibition of proteasome pathway by lactacystin resulted in accumulation of ubiquitylated forms of gelsolin protein.In PANC-1 cell line,there was no significant changes of gelsolin after treatment with lactacystin.Conclusion Ubiquitin-proteasome dependent degradation may be an important regulatory mechanism for gelsolin down-regulation in pancreatic cancer cells.

Objective To investigate the feasibility, efficacy, safety and economy of secondary splenic pedicle trisection method in removing schistosoma cirrhosis caused the splenic function. Methods Thirty patients receiving spleen secondary structure amputation between July 2014 and September 2016 were analyzed. Results Laparoscopic splenectomy with secondary splenic pedicle transaction was successfully performed in 28 patients, whereas two Endo-GIAs were used in 2 patients. The average of operation time was (80 ± 20) min, and operative blood loss was (320 ± 10) ml. The drainage of the splenic fossa was removed (3- 4) days after operation.Postoperative hospital stay was (10.8 ± 1.2) days after operaions. No massive hemorrhage, pancreatic leakage, secondary infection, serious complications such as abscess under diaphragm and recent complication such as infection of incision occurred postoperatively. Platelet of all patients recovered in 4 days postoperatively, and patients with platelet>400 × 109/L was given oral aspirin enteric-coated metformin hydrochloride. All patients were followed up for 6 months postoperatively, and no intestinal obstruction, portal vein thrombosis and other long-term complications occurred in all patients. Conclusions The amputation of secondary structures of the spleen in laparoscopic splenectomy to remove schistosoma cirrhosis caused the splenic function is safe. It could shorten the length of hospital stay and reduce the medical cost. It is a valuable method for clinical promotion.

Une new flavonoids named as notabilisin K (1), together with four known compounds, morusin (2), mulberrofuran A (3), neocyclomorusin (4) and mornigrol F (5) are separated from 95% ethanol extracts of the twigs of Morus notabilis. Compounds 2-5 are separated from this plant for the first time. Notabilisin I, notabilisin J exhibits certain effect against cells of HCT-116, HepG2 and A2780 with IC50 values ranging from 1.47 μmol x L(-1) to 5.46 μmol x L(-1). Morusin exhibits strong effect against five kinds of human cancer cells (BGC823, A2780, HCT-116, HepG2 and NCI-H1650) with IC50 values ranging from 0.74 μmol x L(-1) to 1.58 μmol x L(-1).

Objective To evaluate the endoscopic and pathologic characteristics and etiological ex-amination of viral esophagitis. Methods The data of 16 patients with viral esophagitis, including endoscop-ic, pathological and immunohistochemical findings were retrospectively studied. Results Endoscopic find-ings of viral esophagitis were characterized by single or multiple round and oval ulcers, located at the upper and middle esophagus. The surface of the ulcer was clean, and the boundary was distinct. Pathologic findings included degeneration and necrosis in squamons epithelium, accompanied by ulcer, infiltration of neutrophils and lymphocytes, hyperplasia of capillaries and basal cells and formation of granulation tissues, Immunocyto-chemical examination showed HSV-1 was positive in biopses, while CMV, EBV, HHV8 were negetive. Con-dusion Viral esophagtitis exhibited distinctive endosoopic and pathological features, and etiology can be confirmed by immunohistochemical examinations.

Objective To assess the clinical value of endoscopic uhrasonography(EUS)combined with the mini-probe endoscopic uhrasonography(MPUS)in determing tumor invasion depth and lymph node metastases of early superficial esophageal cancer.Methods One hundred and twenty-four superficial esophageal cancer lesions of 121 patients were staged by EUS combined with MPUS,and the results were finally compared with pathological findings of surgical specimens or samples obtained by mucosal resection.Results The diagnostic accuracy of EUS in T staging of superficial esophageal cancer was 82.3%(102/124).The total ratio of lymph node metastases was 5.0%(6/121),with no node metastases in carcinoma in situ,1.3%(1/28)in mucosal carcinoma,11.6%(5/43)in submucosal carcinoma.Conclusion EUS combined with MPUS is accurate in staging of the superficial carcinoma,which can help the choice of therapeutic strategies.