Objective To explore the role of metabotropic glutamate receptors (mGluRs)group Ⅲ and its subtypes mGluR7 and mGluR8 in nucleus tractus solitarius(NTS)in cardiac-somatic motor reflex (CMR),and to clarify the modulation role of mGluR Ⅲ and its subtypes in NTS in cardiac nociceptoion.Methods 40 SD rats were randomly divided into L-AP4 group,microinjection of mGluRs Ⅲ agonist L-AP4 0.1,1.0,10.0 or 20.0 nmol in NTS;AMN082 group,microinjection of mGluR7 agonist AMN082 1,2 or 4 nmol;DCPG group,microinjection of mGluR8 agonist DCPG 4, 6 or 8 nmol;MSOP group, microinjection of mGluR Ⅲ antagonist MSOP 20 or 100 nmol,20 nmol MSOP+410 nmol L-AP,20 nmol MSOP+2 nmol AMN082,20 nmol MSOP+6 nmol DCPG. The changes of CMR of the rats in various groups were observed.Results Compared with control,the CMR in L-AP4 and AMN082 groups was decreased (P<0.05);the CMR in DCPG group was increased (P<0.05);the CMR in MSOP group after injection of 20 nmol MSOP had no change (P>0.05);the CMR in MSOP group after injection of 100 nmol MSOP was increased (P<0.05);the CMR in MSOP group after injection of 20 nmol MSOP followed L-AP4 or AMNO82 had no change (P>0.05).Conclusion The group Ⅲ mGluRs in the NTS play an inhibitory role in cardiac nociception, and mGluR7 has anti-nociceptive effects while mGluR8 has pro-nociceptive effects.

Objective To study the role of glutamate receptor subtypes in nucleus tractus solitarius(NTS)in cardiac-somatic motor reflex (CMR)induced by intrapericardial administration of capsaicin,and to clarify the modulation mechanism of NTS to cardiac nociceptoion.Methods 60 SD rats were randomly divided into ibotenic (IBO)group, glutamate group, MK-801 group, MCGP group, MK-801 + MCPG group and DNQX group. The NTS microinjected with 130 mmol·L-1 IBO 100 nL,100,200,500 mmol·L-1 L-glutamate 100 nL,NMDA receptor antagonist 40 and 60 mmol · L-1 MK-801 100 nL, metabotropic glutamate receptors antagonist 25 and 50 mmol·L-1 MCPG 100 nL,25 mmol· L-1 MCPG 50 nL plus 40 mmol· L-1 MK-801 50 nL,non-NMDA receptor antagonist 20 and 50 mmol·L-1 DNQX 100 nL,respectively.The changes of CMR of the rats in various groups were observed.Results Compared with control group,the CMR of the rats in IBO group was decreased (P<0.05);the CMR was increased as the concentration increased in glutamate group(P<0.05);the CMR in MK-801 and MCPG groups were decreased (P<0.05);the CMR in MK-801+MCPG group was decreased (P<0.05);the CMR in DXQX group had no changes (P>0.05).Conclusion NTS play an facilictory role in cardiac nociception,and the NMDA receptors and mGluRs receptors mediate this facilitory modulation.

Objective To investigate the expression of Hedgehog signaling pathway members,Shh,Gli1,Sufu and TAK1 as well as phosphorylation-TAK1 (p-TAK1) protein in human pancreatic carcinoma tissues and to explore its relationship between the expression of proteins and the clinicopathologic parameters.Methods The expressions of Shh,Glil,Sufu,TAK1,p-TAK1 proteins in 38 samples of pancreatic cancer tissues and pairing adjacent normal pancreatic tissues were detected by immunohistochemical method.The relationship among the proteins and the relationship between the expression of the proteins and the clinicopathologic parameters were examined.Results The positive expression rates of Shh,Glil,Sufu,TAK1,p-TAK1 protein in human pancreatic carcinoma were 86.8％ (33/38),52.6％ (22/38),68.4％(26/38),55.3％ (21/38),52.6％ (20/38),but they were not expressed in adjacent normal pancreatic tissues.Gli1 expression was positively related to distant metastasis and clinical staging (r =0.524,0.361,P ＜0.05),Sufu expression was positively related to patient's gender (r =-0.378,P ＜0.05),TAK1 expression was positively related to clinical staging (r =0.468,P ＜ 0.05),p-TAK1 expression was positively related to clinical staging and distant metastasis (r =0.418,0.361,P ＜ 0.05).Gli1 expression was positively related to TAK1 and p-TAK1 expression in pancreatic cancer tissues (P ＜ 0.05).Conclusions Hedgehog signaling pathway and p-TAK1 may play a role in the pathogenesis of pancreatic cancer,and the two pathways may interact with each other.

Objective To investigate Toll-like receptor-4 (TLR4) protein expression in human pancreatic adenocarcinoma,and to evaluate the relationship between TLR4 protein expression and angiogenesis.Methods Sixty-two surgically resected human pancreatic adenocarcinoma specimens and 35 normal para-cancerous tissues were investigated for TLR4 protein expression by immunohistochemical SP methods,and CD31 antibody was used to mark microvascular endothelial cells and determine the microvessel density (MVD).The correlation among TLR4 protein expression and MVD and clinicopathologic features of pancreatic adenocarcinoma were analyzed.Results TLR4 protein positive expression rate and MVD in human pancreatic adenocarcinoma was 74.2％ (46/62) and 47.3 ± 13.5,respectively,which were significantly higher than those in the normal pancreatic tissue [17.1％ (6/35),12.6 ±4.8; P ＜0.01].TLR4 protein positive expression rate in the cases with lymph node metastasis was 83.8％,which was significantly higher than that in the cases without lymph node metastasis (60.0％,P =0.036).TLR4 protein positive expression rate in the patients with stage Ⅲ and Ⅳ of TNM classification was 85.3％,which was significantly higher than that in the patients with stage Ⅰ and Ⅱ (60.7％,P=0.028).MVD was closely related to tumor size,lymph node metastasis and TNM stage of pancreatic adenocarcinoma (P =0.008,0.036,0.010).There was a strong positive correlation between TLR4 protein expression and MVD (r =0.534,P ＜0.01 ).Conclusions TLR4 protein expression is closely related to the development and progression of human pancreatic adenocarcinoma and its potential mechanism is related to the promotion of tumor angiogenesis.

Objective To observe the effect of triptolide combined with gemcitabine on proliferation and apoptosis of pancreatic cancer cells ,and to analyze the relevant mechanisms .Methods After treated with TPL ,GEM or TPL combined with GEM in vitro , PANC-1 cells proliferation was accessed by MTT assay and the interaction between the two drugs was calculated .Apoptotic mor-phological changes and apoptosis rate of the cells were investigated by Hoechst 33258 staining and flow cytometry ,respectively .The expression of signal transduction and transcription factor 3(STAT3) ,cysteine aspartate specific proteases-3(caspase-3) protein were detected by Western blot analysis .Results TPL ,GEM or TPL combined with GEM could significantly inhibit the prolifera-tion of PANC-1 cells ,and the combination of the two drugs had a synergistic effect .The cells of the TPL group ,GEM group ,as well as the combined group showed typical apoptotic morphological changes .Compared with the TPL group and GEM group ,the number of apoptotic cells of the combined group increased significantly .Compared with the control group ,the cells apoptosis rate of the TPL group ,GEM group and combined group was significantly increased (P< 0 .05) ,and the apoptosis rate of the the combined group was significantly higher than that of the monotherapy group(P<0 .05) .TPL combined with GEM synergistically inhibited p-STAT3 protein expression and activated caspase-3 protein expression .Conclusion TPL combined with GEM can synergistically in-hibit proliferation and induce apoptosis of pancreatic cancer PANC-1 cells ,its mechanism is related to the inhibition of STAT 3 sig-naling pathway ,promotion the expression of caspase-3 protein .

Objective To investigate the expression of peroxisome proliferator-activated receptor-γ(PPAR-γ) and angiotensin receptor (AT2R) in chronic pancreatitis tissue. Methods Between April 2006 and February 2009, 24 patients were pathologically diagnosed as chronic pancreatitis were enrolled and 12 samples of normal pancreatic tissues were treated as controls. Immunohistochemical staining was used to detect PPAR-γ and AT1R, AT2R expression in pancreatic tissues. Results PPAR-γ and AT1R were mostly negatively expressed in normal pancreatic tissues, the positive scores were 0.33±0.49 and 0.42 ± 0. 51, respectively, while AT2R were weakly positively or negatively expressed in acinus and islet cell, and it was weakly positively expressed in ductal epithelial cells, the positive score was 2. 33 ± 1. 37. In chronic pancreatitis tissue, PPAR-γ, AT1R and AT2R were positively expressed in acinus, ductal epithelial cells, and islet cell, the positive scores were 3.28 ± 2.46, 4.36 ± 2.80 and 4.61 ± 2.89, which were significantly higher than those in the normal group (P<0.01, P <0.01, P<0.05). Conclusions PPAR-γ, AT1R and AT2R were positively expressed in chronic pancreatitis tissue, and they may be the treatment target of chronic pancreatitis.

Objective To assess the safety of endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA)of pancreatic lesions.Methods Patients who underwent EUS-FNA of a pancreatic lesion between January 2005 and June 2007were studied retrospectively.Possible risk factors were assessed by using logistic analysis.Results In 119 patients who underwent pancreatic EUS-FNA,mild acute pancreatitis were observed in 1(0.84%)patient after the operation.No complication occurred in 12 patients with regional portal vein hypertension.Nine patients(7.6%)showed hyperamylasemia 3 h after the procedure,rangeing from 197 to 835 U/L,with an average of(327±200)U/L.Blood amylase level kept increasing 24 h postoperatively in 6 cases of the 9.Logistic regression analysis showed past history of acute pancreatitis,gender,needle size,number of puncture,cystic foci,preoperative blood amylase level and location of foci would not possibly be the risk factors of hyperamylasemia.Conclusion Incidence of complications after EUS-FNA is 0.84%,and the occurrence rate of hyperamylasemia is 7.6%,indicating,EUS-FNA is a safe procedure.

Objective To investigate the expression of Fascin mRNA and its protein in human pancreatic carcinoma cell line and pancreatic cancer tissues and to explore the relationship between the expression of Fascin protein and the clinicopathologic parameters. Methods The expression of Fascin mRNA in pancreatic cancer cell lines (SW1990, Patu8988, BxPC3, cfPAC1) were measured by RT-PCR and the expression of Fascin protein in 54 samples of pancreatic career tissues and 42 adjacent normal pancreatic tissues was detected by immunohistochemical method. Results The expression of Fascin mRNA was confirmed in 3 of 4 pancreatic cancer cell lines (SW1990, Patu8988, cfPAC1), but not in the cell line of BxPC3; the rate of positive expression of Fascin protein in pancreatic cancer tissues was 64.81% (35/54) and there was no positive expression in adjacent normal pancreas tissues; the expression of Fascin protein correlated with the differentiation degree (P < 0.01) and with the lymphatic metastasis of the carcinoma (P <0.05), but not with the size and distant metastasis of the carcinoma (P > 0. 05). Conclusions Fascin protein was highly positively expressed in pancreatic cancer tissues, and the expression of Fascin protein may help diagnose pancreatic, carcinoma and predict the malignant degree.

Objective To investigate the effect of needle sizes and aspiration techniques on sample quantity. Methods Aspiration was performed on porcine liver in vitro for 10 times with three different sizes of needles(19 G, 22 G and 25 G) and four different aspiration techniques[non?negative pressure(NP), 10 ml NP,20 ml NP and slow?pull], 20 mm in depth. A total of six aspirations were performed with each needle by the same aspiration technique. All the obtained specimens were fixed in formalin with the cell block method. The samples were evacuated according to our grading criteria. Results The mean±standard deviation(SD) score for 19 G,22 G, 25 G were 5?71±0?69,4?63±1?24, 3?79±1?84 respectively. The mean±SD score for methods non?NP,10 ml NP,20 ml NP and slow?pull were 4?72±1?53,4?56±1?46,4?72±1?50,4?83±1?76 respectively. The multi?analysis of variance results showed that there were statistical differences between different needles size( F=12?00,P<0?001) with 19 G being the best,followed by 22 G and the least specimen obtained by 25 G needle. There were no statistical differences among aspiration techniques ( F=0?128, P=0?943).The analysis showed that the thicker the needle was,the better sample quality was 19 G yielded to the highest quantity of specimens. The most specimens could be obtained with 19 G needle and non?NP, 22 G needle and 20 ml NP and 25 G needle and slow?pull. Conclusion In clinic, aspiration technique should be selected according to different aspiration needles. 19 G is superior to others, with non?NP method. For 22 G needle, 20 ml NP is preferred and for 25 G needle,slow?pull is preferred.

Objective To investigate the expression of TGF-β activated kinase1 (TAK1),phosphoryted TAK1 (p-TAK1) and the density of tumor associated macrophages (TAM) in human pancreatic carcinoma tissues and pairing adjacent normal pancreatic tissues,and explore their relationship.Methods The expression of TAK1,p-TAK1,CD68 (the marker of tumor associated macrophages) proteins in 57 samples of pancreatic cancer tissues and 35 samples of pairing adjacent normal pancreatic tissues were detected by immunohistochemical method,then the density of TAM was evaluated.The relationship between the protein expression and TAM,and the relationship between them and clinicopathologic parameters were examined using SPSS 18.0 software,and the independent risk factors of TNM staging were analyzed by multivariate logistic regression.Results TAK1 and p-TAK1 were positively expressed in 42.1％ (24/57) and 40.4％ (23/57) pancreatic carcinoma tissues,significantly higher than adjacent normal pancreatic tissues [14.3 ％ (5/35) and 11.4％ (4/35)];the proportion of pancreatic carcinoma tissues with high density TAM was 38.6％ (22/57),higher than that of adjacent pancreatic tissues [8.6％ (3/35)],the differences were statistically significant (P＜0.05).TAK1 expression was positively related to tumor size,tumor differentiation,lymph node metastasis,distant metastasis and clinical staging;p-TAK1 expression was positively related to tumor differentiation,lymph node metastasis,distant metastasis and clinical staging;the density of TAM was positively related tumor differentiation,lymph node metastasis,distant metastasis and clinical staging (all the P values were less than 0.05).The expression of TAK1 and p-TAK1were positively correlated with the density of TAM (P ＜0.001).Multivariate logistic regression analysis showed that high density of TAM was independently associated with advanced clinical staging (P =0.002,OR =129.5,95％ CI 6.2 ～2718.6).Conclusions TAK1 pathway and TAM may play an important role in the pathogenesis and progression of pancreatic cancer,and there may be synergy effect between them.