Objective To study the role of Caspase-1 activated cytokines in acute lung injury (ALI) in experimental severe acute pancreatitis (SAP). Methods A rat model of SAP was induced by retrograde infusion of 5% sodium taurocholate into the biliary pancreatic duct. Thirty-two SD rats were randomly divided into four groups: healthy control (HC), SAP 6h, SAP 12h and SAP 18h groups. Serum IL-1? level was measured by ELISA. Intrapulmonary expressions of Caspase-1, IL-1? and IL-18 mRNA were detected by semi-quantitative RT-PCR. Results The serum IL-1? levels were significantly increased after the induction of pancreatitis (P

Objective The core mechanism of renal insterstitial fibrosis (RIF) is epithelial-mesenchymal transition.This study aimed to investigate the effect of erythropoietin on high glucose-induced epithelial-mesenchymal transition ( EMT) of normal hu-man kidney proximal tubular epithelial cells (HK-2) and its possible mechanism. Mothods HK-2 cells cultured in vitro were ran-domly divided into a blank control group , a high glucose induction group , a mannitol induction group , an EPO induction group , an EPO (5, 10, and 20U/mL) inhibition group, and an Rho kinase inhibitor group.After 24 hours of intervention, the mRNA levels of RhoA and ROCK were determined by RT-PCR, those of E-cadherin and α-smooth muscle actin (α-SMA) proteins detected by immu-nofluorescence staining , and the expression of FN proteins in the supernatant measured by ELISA . Results Compared with the blank control group , the expressions of RhoA and ROCK 1 mRNA were significantly increased in the high glucose induction group (0.945 ±0.132 vs 1.400 ±0.022, 1.007 ±0.002 vs 1.913 ±0.011, P<0.05), but markedly decreased in the 5, 10, and 20U/mL EPO inhibition groups (1.400 ±0.022 vs 1.278 ±0.006, 1.400 ±0.022 vs 0.770 ±0.005, 1.400 ±0.022 vs 0.334 ±0.009, P<0.006) in comparison with the high glucose induction group , and the effects were related to the concentration of EPO .Compared with the blank control, the expression of E-cadherin protein was increased in the high glucose induction group (0.644 ±0.006 vs 0.107 ± 0.004, P<0.05), but remarkably decreased in the 5, 10, and 20 U/mL EPO inhibition groups (0.236 ±0.006, 0.433 ±0.010, 0.521 ±0.010) in comparison with the high glucose induction group (P<0.05), and the effects were also related to the concentration of EPO.Pearson correlation analysis showed a positive correlation between the mRNA expressions of RhoA and ROCK 1 in the high glu-cose induction and EPO inhibition groups . Conclusion EPO can inhibit high glucose-induced epithelial-mesenchymal transition of normal human kidney HK-2 cells and thus delay renal fibrosis , which mignt be related to the RhoA/ROCK signaling pathway .

23 cases of the patients with supraorbital neuralgia were treated by puncturing Yangbai (GB 14) toward Yuyao (Extra), Zanzhu (BL 2), Taiyang (Extra), Touwei (ST 8),Zhongzhu (TE 3) and Neiting (ST 44) on the sick side, plus laser radiation on a site about 1 cm apart from the midpoint of the eyebrow of the sick side. After 10 treatments, the results showed cure in 19 cases and remarkable effect in 4 cases.

Objective To explore the application value of preoperative methylene blue staining in locating for the operation of intraspi-nal tumors. Methods The clinical data of patients with intraspinal tumors from September 2010 to September 2012 in our hospital were ret-rospectively analyzed. The patients were divided into tag group and control group according to whether stained by methylene blue or not. The operation time( min) ,intraoperative hemorrhage,the rate of total resection of tumor,spinal instability rate,tumor recurrence rate,and reopera-tion rate of two groups were compared. Results The operation time of tag group was significantly shorter than that of the control group. The amount of intraoperative bleeding was significantly less than that of in control group, the differences were statistically significant(P<0. 05). The total resection rate of tumor was significantly higher than that in control group,the differences were statistically significant(P<0. 05). The spinal instability rate,tumor recurrence rate and operation rate of patients within 1 year in two groups were not significant. Conclusion The methylene blue method is simple and convenient,and provides favorable conditions for the operation,which reduces the operation time and intraoperative hemorrhage,increases the rate of complete tumor resection. There was no difference in recurrence rate,operation rate and the stability of the spine within 1 year compared to traditional method.

Objective Clinical treatment can delay the development of renal interstitial fibrosis , but it can not reverse renal dysfuntion.The article was to discuss the influence of recombinant human erythropoietin ( rHuEPO ) on inflammatory factors in the process of renal interstitial fibrosis and its possible mechanism . Methods The vitro cultured HK-2 cells were randomized into 7 groups:the blank control group , rHuEPO control group ( addition of 20U/mL rHuEPO), albumin stimulation group (addition of 5mg/mL albumin), 5mg/mL rHuEPO intervention group (5mg/mL albumin +5U/mL rHuEPO), 10 U/mL rHuEPO intervention group (5mg/mL albumin +10 U/mL rHuEPO), 20U/mL rHuEPO intervention group (5mg/mL albumin +20U/mL rHuEPO), and Rho inhibi-taion group (addition of 5mg/mL albumin 30min after 10μmol/L Y27632), 24 h acting time for each group.We observed the changes of cell morphology in each group .Reverse transcription polymerase chain reaction ( RT-PCR) was used to evaluate the mRNA levels of RhoA, ROCK1 and IL-6 , and ELISA was applied to measure the levels of supernatant TNF-αand IL-6 protein. Results The form of pebbles or paving stone was observed in blank control group and rHuEPO intervention groups , a long and thin spindle change with the appearance of fibre cells in albumin stimulation group , the transformation to pebbles in 5, 10, 20 mg/mL rHuEPO intervention groups , the form of oval and slightly increased intercellular space in Rho inhibitaion group .Compared with the blank control group , the expressions of RhoA mRNA, ROCK1 mRNA and IL-6 mRNA significantly increased in the albumin stimulation group (P<0.05), while significantly reduced in 5, 10, 20 mg/mL rHuEPO intervention groups (P<0.05), which was in negative relation with the rHuEPO concentrations .Compared with the albumin stimulation group , the expressions of ROCK 1 mRNA and IL-6 mRNA reduced in Rho inhibtation group (P<0.05), while there was no significant difference as to the expression of RhoA mRNA .ELISA results showed:compared with blank control group , the expressions of supernatant TNF-α([452.32 ±33.23] ng/L vs [1347.54 ±41.52] ng/L), IL-6 protein([884.62 ±0.73] pg/L vs [95.12 ±0.32]pg/LP<0.05) increased significantly.Compared with albumin stim-ulation group, the expressions of TNF-αin 5, 10, 20 mg/mL rHuEPO intervention groups and Rho inhibitation group reduced signifi-cantly([1003.32 ±3.42] ng/L, [821.32 ±21.32] ng/L, [590.15 ±7.68] ng/L, [488.13 ±65.03] ng/L vs [1 347.54 ± 41.52]ng/L,P<0.05), while the expressions of IL-6 mRNA reduced accordingly in 5, 10, 20 mg/mL rHuEPO intervention groups and Rho inhibitation group reduced significantly ([656.68 ±0.55] pg/L, [422.35 ±0.22] pg/L, [217.32 ±0.35] pg/L, [309.49 ±0.21] pg/L vs [884.62 ±0.73]pg/L,P<0.05).Moreover, there was significant statistical difference among 5, 10, 20 mg/mL rHuEPO intervention groups(P<0.05). Conclusion RHuEPO can inhibit the transdifferentiation process of HK-2 cells in-duced by albumin by suppressing inflammation factors , and the mechanism may be involved in RhoA/ROCK signaling pathway .

Objective To investigate the relationship between EphA2, EphrinAl and E-cadherin expressions and tumor stage and prognosis in pancreatic cancer. Method EphA2, EphrinAl and Ecadherin expressions were detected by immunohistochemistry in the tumor tissue and normal tissue specimens from 48 patients with primary pancreatic cancer. Results The expressions of EphA2 and EphrinAl were higher in the pancreatic carcinoma tissues than in the normal pancreatic tissues (P<0. 05). The E-cadherin expression was lower in the pancreatic cancer tissues than in the normal pancreatic tissues (P<0. 05). With decreasing histological differentiation, the expressions of EphA2 and EphrinAl in carcinoma tissues increased significantly (P<0. 05), while the E-cadherin expression decreased significantly (P<0. 05). The positive expressions of EphA2 and EphrinAl in the primary tumor significantly increased in stageⅢ and Ⅳ than in stage Ⅰ and Ⅱ (47. 9%vs 6. 25% , P<0. 05;47. 9% vs 8. 3%, P<0. 05), while the negative expression of E-cadherin was reversely correlated with these tumor stages (14. 6% vs 64. 6%, P<0. 05). Cox multivariate analysis showed that the clinical stage, EphA2 positive expression and E-cadherin negative expression were significantly associated with survival. Conclusion Abnormal expressions of EphA2, EphrinAl and E-cadherin were involved in the progression of pancreatic cancer and they were useful in predicting prognosis.

Objective To investigate the changes of extraceUular regulated kinase 1/2 (ERK1/2) phosphorylation and assess the effects of blocking the ERK1/2 phosphorylation on rats with acute necrotizing pancreatitis (ANP). Methods The ANP model was induced by retrograde injection of 5% sodium tanrocholate into the biliary and pancreatic duct. 5 rats were treated as normal control. Other 75 Sprague-Dawley (SD) rots were randomly divided into sham operations(SO) group (n =25), ANP group (n =25) and PD98059 group (n =25). The rats were sacrificed at 15 min, 30 min, 1 h, 3 h and 6 h after ANP induction, the blood and pancreatic sample were taken. Pathological changes of pancreas were observed with light microscope and scored. The serum level of TNF-α and IL-1β was determined by ELISA. MPO activities in pancreas were measured by enzyme chemistry assay. Western blotting was performed to determine the phosphorylations of ERK1/2 in the pancreas homogenates. Results There was no significant pathologic changes in rats of SO group;but significant injuries occurred in ANP group, the pathologic score at 3 h was 9.9 ± 0.4;the extent of injuries attenuated in PD98059 group, the pathologic score at 3 h was 4.0 ± 0.4 (P < 0.05). The serum levels of TNF-α at 3 h in SO, ANP and PD98059 groups were (65.8 ± 20.5) pg/ml, (286.5 ± 50.3) pg/ml, (180.4±32.9)pg/ml, respectively;the serum levels of IL-1β at 3 h in SO, ANP and PD98059groups were (85.8 ± 25.5) pg/ml, (293.8 ± 46.3) pg/ml, (200. 5 ± 33.6) pg/ml, respectively;MPOactivities in pancreas were (0. 19 ± 0.02)U/g, (0.61±0.05)U/g, (0.52±0.03) U/g, and the values in ANP and PD98059 groups were significantly higher than those in SO group, while the values in PD98059 group were significantly lower than those in ANP group (P < 0.01). The expression of ERK1/2 phosphorylation in normal pancreas was 1100 ± 141, the expressions of ERK1/2 phosphorylation in ANP group at 15 min, 30 min were 5300 ± 486, 5621 ± 384, respectively;the expressions began to decrease 1 h later and returned the similar level as SO group at 6 h;the expressions of ERK1/2 phosphorylation in PD98059 group at 15 min, 30 rain were 4200 ± 370, 3600 ± 290, respectively;which were signifieanfly lower than those in ANP group (all P value < 0. 01). Conclusions The ERK1/2 signal transduetion pathway plays an important role in the pathogenesis of ANP. Inhibition of ERK1/2 phosphorylation by PD98059 may decrease the production of IL-1β, TNF-α and pancreatic MPO, attenuate the extent of pancreatic pathologic injuries.

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 observe the effect of Codonopsis and Glycyrrhiza glycoconjugates on migration and membrane potential of small intestinal epithelial cells(IEC-6),and to explore the promoting effects of Yiqi jianpi herb Codonopsis and Glycyrrhiza on gastrointestinal mucosal injury repair and the underlying mechanisms. METHODS Under normal conditions or loaded with the inhibitor of potassium channel 4-aminopyridine(4-AP),IEC-6 cells were treated with Codonopsis and Glycyrrhiza glycoconjugates (25-200 mg · L-1) for 24 h,respectively. IEC-6 cell migration was observed by the phase contrast microscope and cell membrane potential was detected by flow cytometry. RESULTS Codonopsis and Glycyrrhiza glycoconjugates (50 and 100 mg · L-1) increased the number of migrated IEC-6 cell compared with normal control group(P<0.01,P<0.05). Compared with normal control group,4-AP reduced the number of migrated IEC-6 cell(P<0.01). Codonopsis and Glycyrrhiza glycoconjugates (50-200 mg · L-1)reversed cell migration inhibited by 4-AP significantly when compared with 4-AP model group(P<0.01). The results of flow cyometry analysis showed that the cell membrane poten?tial was increased after treatment with Codonopsis and Glycyrrhiza glycoconjugates(50 mg · L-1)compared with normal control group and resulted in an increase in cell membrane hyperpolarization(P<0.01). Compared with normal control group,4-AP decreased the cell membrane potential(P<0.01)and resulted in cell membrane depolarization. Also,compared with 4-AP model group,cell membrane depolarization induced by 4-AP was reversed by treatment with Codonopsis and Glycyrrhiza glycoconjugates(100 and 200 mg·L-1). CONCLUSION Codonopsis and Glycyrrhiza glycoconjugates may promote gastrointestinal mucosal injury repair and the mechanisms may involve the activation of signaling pathways by affecting polyamine-dependent intestinal epithelial cell migration voltage-gated K+channels.

Objective To study the effects of erythropoietin (rhEPO) in high glucose induced proliferation and apopto?sis of human kidney proximal tubular epithelial (HK-2) cells, and the possible mechanism thereof. Methods HK-2 cells cultured in vitro were divided into several groups randomly:blank control group, high glucose group, mannitol group, rhEPO control group, different concentrations of rhEPO treatment groups (5, 10, 20 U/mL) and Rho kinase group. The reverse tran?scription polymerase chain reaction (RT-PCR) was used to evaluate the mRNA levels of RhoA and ROCK after 24 hours. Tetrazolium salt method (MTT) was used to determine the cell proliferation. Cell apoptosis was detected by flow cytometry. Results Compared with blank control group the expression levels of RhoA and ROCK1 mRNA were significantly in?creased in high glucose group (P < 0.05). RhoA, ROCK1 mRNA expressions significantly decreased in rhEPO group than those of high glucose group (P<0.05). There was a positive correlation between the expression levels of RhoA mRNA and ROCK1 mRNA in high glucose group and rhEPO group. MTT method showed that rhEPO significantly promoted the prolifer?ation of HK-2 cells (P<0.05). Flow cytometry analysis showed that high glucose induced apoptosis in HK-2 cells, which was significantly inhibited in rhEPO group and Rho kinase group as compared to that of high glucose group in a concentra?tion dependent manner (P<0.05). Conclusion rhEPO can promote HK-2 cell proliferation and inhibit apoptosis, which may be related to RhoA/ROCK signaling pathway.