1.ROLE OF CASPASE-1 ACTIVATED CYTOKINES IN ACUTE LUNG INJURY IN EXPERIMENTAL SEVERE ACUTE PANCREATITIS
Xiaohua ZHANG ; Zhaoshen LI ; Zhenxing TU
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
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
2.Inhibitory effect of erythropoietin on high glucose-induced transition of human kidney proximal tubular epithelial cells and its possible mechanism
Yanxia CHEN ; Xiaohua QIN ; Xiangdong FANG ; Chong HUANG ; Weiping TU
Journal of Medical Postgraduates 2014;(10):1038-1041
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 .
3.Treatment of 23 Cases of Supraorbital Neuralgia by Acupuncture plus Semi-conductive Laser
Xiaohua TU ; Luoyan YANG ; Zengli LI ; Guoqi HUANG
Journal of Acupuncture and Tuina Science 2007;5(2):113-114
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.
4.Role of extracellular regulated kinase signal transduction pathway in the pathogenesis of acute necrotizing pancreatitis
Xingang SHI ; Yongchun XU ; Xiaohua MAN ; Jing JIN ; Yanfang GONG ; Zhenxing TU ; Zhaoshen LI
Chinese Journal of Pancreatology 2010;10(2):116-119
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.
5.Expression and significant of EphA2、 EphrinAl and E-cadherin in pancreatic carcinoma
Feng LU ; Zurong YUAN ; Jianxiong TANG ; Wei WANG ; Xiaohua SONG ; Yanyuan TU ; Wenhao TANG
Chinese Journal of Hepatobiliary Surgery 2011;17(6):471-474
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.
6.The expression and significance of Fascin mRNA and its protein in pancreatic carcinoma cell line and pancreatic cancer tissues
Qingyun ZHU ; Zhaoshen LI ; Shude LI ; Xiangui HU ; Xiaohua MAN ; Zhenxing TU
Chinese Journal of Pancreatology 2008;8(5):305-307
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.
7.Application of methylene blue staining in operation of intraspinal tumors
Jiaqi LIAO ; Jinxian XU ; Yong TU ; Xiaohua WEN ; Hanwen LIU ; Feng LI
Journal of Regional Anatomy and Operative Surgery 2013;(6):597-599
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.
8.Clinical observation of partial pancreatectomy as part of primary cytoreductive surgery in advanced epithelial ovarian cancer
Libing XIANG ; Yunxia TU ; Tiancong HE ; Xuan PEI ; Xuxia SHEN ; Wentao YANG ; Xiaohua WU ; Huijuan YANG
Chinese Journal of Obstetrics and Gynecology 2016;51(5):361-365
Objective The aim of this study is to evaluate the safety and efficacy of partial pancreatectomy as part of primary cytoreductive surgery in advanced epithelial ovarian cancer (EOC). Methods A total of 8 patients were recruited in this study who underwent partial pancreatectomy during the primary cytoreductive surgeries for advanced EOC in Fudan University Shanghai Cancer Center from April 2009 to July 2015. Their clinicopathological characteristics, diameter of metastatic tumors, the scope of cytoreductive surgeries, residual diseases after cytoreductive surgeries, postoperative complications and survival situation were retrospective analyzed. Results (1) Clinicopathological characteristics:the median age of these patients was 58 years old(range: 39-63 years old). The median value of preoperative serum CA125 was 1 688 kU/L(range: 119-5 000 kU/L). The median diameter of metastatic tumors involved in pancreatic body or tail was 4.5 cm (range:3-10 cm). All the tumors from the 8 patients were confirmed to be high-grade serous carcinoma. Four patients were staged as International Federation of Gynecology and Obstetrics (FIGO)Ⅳ, and the other 4 patients were staged as FIGOⅢc. (2) Tumor metastases and the scope of cytoreductive surgeries:all of these 8 patients had widely disseminated ovarian cancer, with involvement of upper abdominal, middle abdominal and pelvic cavity. Each patient underwent extensive intra-abdominal cytoreductive surgeries, including hysterectomy, bilateral salpingo-oophorectomy, omentectomy, pelvic periton-ectomy, splenectomy, partial pancreatectomy. Each patient had cytoreductive surgeries of 9.6 different sites on average. Of all 8 patients who underwent partial pancreatectomy, 7 patients had pancreatic tails removed;the other 1 patient had pancreatic body and tail removed. The median volume of blood loss during surgery was 1 350 ml(range:300-3 500 ml), blood transfusion was performed in 7 patients with the median volume of 1 150 ml (range: 500-1 800 ml). (3) Residual diseases after cytoreductive surgeries: optimal cytoreduction was achieved in all patients, with microscopic residual disease in 3 patients, residual tumors diameter < 0.5 cm in 3 patients, and residual tumors diameter between 0.5 and 1 cm in 2 patients. (4) Postoperative complications: 4 patients suffered from complications including pancreatic leakage (2/8), intraperitoneal hemorrhage (1/8) and pancreatic pseudocyst accompanied by infection (1/8). These complications were treated successfully by conservative managements. (5) Survival situation: during the median follow-up duration of 17 months (ranged from 2 to 46 months), 5 patients were still alive until the end of follow-up, including 4 cases under treatment and 1 case survived 29 months without relapse after treatment. Three patients were respectively died in 5, 20 and 46 months after surgery. Conclusion There is a higher risk of postoperative complications of pancreas resection as part of primary cytoreductive surgery in advanced epithelial ovarian cancer, but the resection of pancreatic metastases and part of the pancreas is feasible and necessary.
9.Effect of Codonopsis and Glycyrrhiza glycoconjugates on migration and membrane potential of IEC-6 cells
Ruliu LI ; Yuzhu TAO ; Peng WEN ; Xiaohua TU ; Jiazhong CAI ; Weiwen CHEN
Chinese Journal of Pharmacology and Toxicology 2015;(6):917-923
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.
10.The influence of rHuEPO on inflammatory factors in the process of renal interstitial fibrosis and its possible mechanism
Luojia JIANG ; Liping YANG ; Xianfeng WU ; Chong HUANG ; Xiaohua QING ; Xiangdong FANG ; Weiping TU
Journal of Medical Postgraduates 2015;(6):594-599
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 .