1.IL-10 reduces Interleukin-1? in rats with acute necrotizing pancreatitis
Dehai DENG ; Zhihai LIANG ; Guodu TANG
Basic & Clinical Medicine 2006;0(11):-
Objective To investigate the effects of recombinant human interleukin(IL)-10 on serum interleukin-1? in L-arginine-induced acute necrotizing pancreatitis of rats.Methods Ninty-two Sprague-Dawley rats were randomly divided into three groups.Group A(n=36) and Group I(n=32) received three intraperitoneal injections of 6% L-arginine(1.0 mg/g) at hourly intervals.Group I(n=32) was treated with 10,000 unites of intraperitioneal recombinant human IL-10 at the 2nd,5th and 8th hour after the last injection of L-arginine.Group C(n=24) received saline alone.Rats were killed at the 4th,12th,24th and 36th hour after the last L-arginine injection.Serum interleukin-1? and amylase were assayed.Pancreatic tissues were examined histopathologically by microscopy.Results Serum amylase,interleukin-1? and pancreatic histopathological scores in group A increased significantly after L-arginine injection(P
2.Study on the Expression of LMP1,CyclinD1 and Bcl-2 in Nasopharyngeal Carcinoma and Its Clinical Significance
Hong ZHU ; Liang ZENG ; Zhihai XIE
Journal of Chinese Physician 2001;0(05):-
Objective To investigate LMP1,CyclinD1 and Bcl-2 expression in nasopharyngeal carcinoma(NPC) and its clinical significance,and analyze their relation. Methods LMP1,CyclinD1 and Bcl-2 expression was assayed respectively in 60 NPC tissues by immunohistochemical method. Results ⑴The positive expression rates of LMP1,CyclinD1 and Bcl-2 in NPC were 65%(39/60), 60%(36/60)and 65%(39/60) respetively; ⑵The mean survival period, the five-year survival rate and median survival period in the NPC patients with LMP1, cyclinD1 and Bcl-2 positive expression were significantly shorter than those in the NPC patients with LMP1, cyclinD1 and Bcl-2 negative expression (P
3.Expression of ghrelin and hormone secretagogue receptor in pancreas of rats with acute necrotizing pancreatitis
Chunyun FANG ; Guodu TANG ; Zhihai LIANG ; Xiping TANG
Chinese Journal of Pancreatology 2012;(6):384-387
Objective To observe the expression of ghrelin and growth hormone secretagogue receptor (GHSR) in pancreas of rats with acute necrotizing pancreatitis (ANP),and investigate the role of ghrelin in the pathogenesis of ANP.Methods Seventy SD rats were randomly divided into ANP group (n =35) and control group (n =35).ANP model was induced by retrograde injection of 4% sodium taurocholate into the biliary and pancreatic duct.Rats in the control group underwent laparotomy with gentle pancreas manipulation only.At 3,6,12,24,48 h after ANP induction,the rats were sacrificed,and the serum level of amylase was determined,pathological changes in pancreatic tissue were routinely observed and scored.The expressions of ghrelin mRNA,protein and GHSR mRNA,protein in pancreas were evaluated by RT-PCR and Western blot.Results Serum amylase level began to increase at 3h after sodium taurocholate injection and reached the peak value at 6 h [(8244 ± 2950) U/L],which was significantly higher than that in control group (P < 0.05).Pancreatic injuries was aggravated with time,the pathologic score at 24 h was (11.91 ± 1.31) score,which was significantly higher than (3.12 ± 1.60) score in the control group.The expressions of ghrelin mRNA and GHSR mRNA in pancreas of ANP group were increased gradually with time,and were significantly higher than those of control group at all time points,at 48 h,1.29 ±0.64 vs 0.58 ±0.05 and 0.94 ±0.16vs 0.19 ±0.03,P < 0.05.The expressions of ghrelin protein and GHSR protein in pancreas of ANP group were significantly higher than that in control group at 12,24,48 h.at 48 h,3.05 ± 0.48 vs 2.18 ± 0.23 and 2.34 ± 0.32 vs 1.55 ± 0.10 (P < 0.05).Conclusions The expressions of ghrelin mRNA,protein and GHSR mRNA,protein of pancreas are significantly increased in rats of ANP,and are associated with the severity of ANP.
4.Effect of lipopolysaccharide on toll like receptor 7 and toll like receptor 9 in AR42J cell lines
Luyi ZHANG ; Guodu TANG ; Xiping TANG ; Zhihai LIANG ; Yuan ZHAN
Chinese Journal of Pancreatology 2014;14(3):163-166
Objective To investigate the roles of toll like receptor7 (TLR7) and toll like receptor 9 (TLR9) in the pathogenesis of acute pancreatitis.Methods AR42J cells were treated by lipopolysaccharide at different dosages (0,1,10,100 mg/L),and cell model of acute pancreatitis in vitro was established.AR42J cells without lipopolysaccharide treatment were as control.Cells and culture supernatant were collected after 24 hours cultivation.TLR7,TLR9 mRNA and protein expressions were detected by RT-PCR and Western Blot,and levels of TNF-α,IL-10 in culture supernatant were measured by ELISA.Results The TLR 7 mRNA expression levels in control group,1,10,100 mg/L lipopolysaccharide group were 0.12 ± 0.09,0.28 ± 0.06,0.49 ± 0.04,0.78 ± 0.04,and the TLR9 mRNA expression levels were 0.06 ± 0.02,0.32 ± 0.03,0.56 ± 0.14,0.84 ± 0.12; the TLR7 protein expression levels were 0.04 ± 0.01,0.26 ± 0.05,0.49 ±0.04,0.77 ±0.16,and the TLR9 protein expression levels were 0.10 ±0.14,0.62 ±0.23,1.21 ± 0.26,1.75 ± 0.13 ; the TNF-α levels in culture supernatant were (8.01 ± 5.32),(25.64 ± 8.71),(49.06 ± 10.23),(75.83 ± 6.65) ng/L,and the IL-10 levels were (155.54 ± 25.47),(105.16 ± 10.49),(69.36 ± 8.19),(14.07 ± 9.06)ng/L.The expression levels of TLR7 and TLR9's mRNA,protein in cell,as well as the levels of TNF-α in culture supernatant increased with the lipopolysaccharide concentration,while the levels of IL-10 in culture supernatant decreased with the lipopolysaccharide concentration,and the difference among these groups was statistically significant (P < 0.01).Conclusions The expressions of TLR7 and TLR9 in AR42J cells treated by using lipolysaccharide are obviously up-regulated,and it suggests that TLR7 and TLR9 may be vital in the pathogenesis of acute pancreatitis.
5.Clinical application of the Classification of acute pancreatitis-2012
Qing WU ; Zhihai LIANG ; Guodu TANG ; Wenjing CHEN ; Chunyun FANG
Chinese Journal of Pancreatology 2013;13(4):217-221
Objective To investigate the clinical value of the Classification of acute pancreatitis2012.Methods Medical records and clinical data of patients with acute pancreatitis (AP) who were admitted to First Affiliated Hospital of Guangxi Medical University between October 2009 and September 2012 were retrospectively reviewed and analyzed.Patients were divided into mild acute pancreatitis (MAP),moderately severe acute pancreatitis (MSAP),and severe acute pancreatitis (SAP) groups according to the Classification of acute pancreatitis-2012.The number of improved and cured patients,length of hospital stay,hospitalization costs,rate of ICU admission,length of ICU stay,incidence of SIRS,and length of SIRS continue,Ranson scores,APACHE Ⅱ scores,computed tomographic severity index (CTSI) scores among the 3 groups were compared.Results One hundred and sixty-six patients with AP (119 males and 47 females) were included,and 76 were MAP,65 MSAP and 25 SAP.The average interval between AP onset and hospital admission was (2.27 ± 1.46) d.The number of improved and cured patients,length of hospital stay,hospitalization costs,rate of ICU admission,length of ICU stay,incidence of SIRS,and length of SIRS continue,Ranson scores,APACHE Ⅱ scores,CTSI scores increased with the severity of AP.The corresponding values in SAP group were 21 cases (84.0%),(23.8 ± 13.6) d,(53900 ± 30260) Yuan,48.0% (12/25) and (5.76 ± 13.8) d,96.0% (24/25) and (5.00 ± 2.40) d,(3.76 ± 1.30) score,(8.52 ± 4.24) score,(5.44 ± 3.48) score.Seventy-nine patients developed local complications,among them 34 was acute peripancreatic fluid collection,45 was acute necrosis collection.The incidence of acute necrosis collection in SAP group was significantly higher than that in MSAP group (68.0% vs 44.6%,P =0.047),but the incidence of acute peripancreatic fluid collection in SAP group was significantly lower than that in MSAP group (16.0% vs 46.2%,P =0.016).Organ failure occurred in 42 patients,among them 35 cases were respiratory failure,2 cases were renal failure,and 5 cases were respiratary and renal failure.The incidence of organ failure in SAP and MSAP group was 100% and 26.2%,the difference between the two groups was statistically significant (P < 0.05).Conclusions Classification of acute pancreatitis-2012 is a simple and convenient system,which can predict the severity of AP and appropriate for clinical application.
6.Expression and role of MLCK in small intestine mucosa in rats with acute necrotizing pancreatitis
Huirong SHI ; Guodu TANG ; Mengbin QIN ; Zhihai LIANG
Chinese Journal of Pancreatology 2015;15(2):106-110
Objective To explore the expression and function of myosin light streptokinase (MLCK) in small intestine mucosa of acute necrotizing pancreatitis (ANP) rats.Methods Fifty-six male SD rats were randomly assigned to control group and ANP group.A rat model of ANP was reproduced by retrograde injection of 4% sodium taurocholate into the biliopancreatic duct,while the control group underwent a sham operation.The rats were sacrificed at 6th,12th,24th,48th hour after ANP induction.Serum amylase、TNF α,IL 1β,diamine oxidase (DAO) were measured.The pathological scores in the pancreas and small intestine were observed.The ultrastructure and tight junction (TJ) changes in the small intestine mucosa were observed with an electron microscope.The localization and expression of MLCK in small intestine mucosa was determined by immunohistochemistry method.Results Compared to the control group,the serum amylase,TNF-α,IL-1 β,DAO level,in the ANP group were all significantly increased;[(4 978 ± 1 574) U/L vs (1 176 ± 124))U/L,(47.88 ± 15.85) μg/L vs (17.24 ± 1.99) μg/L,(132.48 ± 68.54) μg/L vs (23.51 ± 6.44) μg/L,(95.96 ± 30.84)μg/L vs (38.06 ± 17.73)U/L at 12 h],and the pathology scores of pancreas and small intestine were both significantly elevated [12 h:(12.2 ± 1.80) vs (4.68 ± 0.35),(2.58 ± 0.52) vs (0.58 ±0.26)] (P <0.05);the MLCK protein expression in small intestine mucosa was significantly increased in ANP group (12 h:0.1863 ± 0.0230 vs 0.1636 ± 0.0049),and the difference was statistically significant (P <0.05).The small intestine ultrastructure was seriously damaged and TJ was widened significantly in ANP Group.Conclusions The increased serum TNF alpha and IL-1β concentration and DAO activity and up-regulated MLCK protein expression in small intestine mucosa may damage the integrity of tight junction of intestinal epithelial cell and cause intestine mucosa barrier dysfunction.
7.Expression of pancreatic thioredoxin-1 in rats with acute necrotizing pancreatitis and effect of melatonin on thioredoxin-1
Weiyi ZHONG ; Guodu TANG ; Zhihai LIANG ; Mengbin QIN ; Wenfu HUANG ; Jinxian LIANG
Chinese Journal of Pancreatology 2009;9(4):262-265
Objective To investigate the expression of pancreatic thioredoxin-1 (TRX-1) in rats with acute necrotizing pancreatitis (ANP) and the effect of pretreatment of melatonin on its expression. Methods Male Spraque-Dawley rats (n = 12) were randomly divided to ANP group, melatonin group, control group with 24 rats in each group. The rats in ANP group received three intraperitoneal injections of 25 ml/kg body weight 6% L-arginine at an interval of 1 h to induce ANP. The rats in melatonin group received intraperitoneal injections of 25 ml/kg body weight 6% melatonin 30 min before ANP induction; rats in ANP group and control group received intraperitoneal injections of same amount of saline. Rats were sacrificed at 6 h, 12 h and 24 h after ANP induction. The serum level of amylase was measured and the pathological evaluation of pancreatic tissues was performed. The concentrations of malondialdehyde (MDA) and myeloperoxidase (MPO) in pancreatic tissues were measured. The expressions of TRX-1 protein were detected by immunohistochemistry and the expressions of TRX-1 mRNA in pancreatic tissues were determined by RT-PCR.Results In ANP group, serum level of amylase, MDA, MPO, TRX-1 mRNA and TRX-1 protein in pancreatic tissues were (3 012 ±1 425) U/L, (4.13 ± 1. 85)nmol/mg prot,(7.45 ± 1.26)nmol/mg prot, 0.68 ±0. 18, 66.8 ±8. 1, while they were (1 835±499)U/L, (3.03 ±2.12) nmol/mg prot, (5. 32 ± 1.06) nmol/mg prot, 0.50±0.09, 80. 29 ±8. 14, respectively in melatonin group, the values in melatonin group were significantly lower thanthose in ANP group (P < 0.05). The peak value of TRX-1 mRNA and TRX-1 protwein expressions shifted from 12 h after ANP induction in ANP group to 6 h after ANP induction in melatonin group. Conclusions The expression of pancreatic TRX-1 protein and TRX-1 mRNA in rats with ANP was significantly increased. Melatonin pretreatment could promote pancreatic tissues to express TRX-1 protein and TRX-1 mRNA, and may be protective for pancreatic tissues damages.
8.The expression of TLR4 in fiver cell treated with cyst fluid of Echinocaccus granulosus
Yalou ZHANG ; He HUANG ; Quanhua FANG ; Tao LIU ; Zhihai PEN ; Liang LI ; Hao WEN
Chinese Journal of Microbiology and Immunology 2009;29(9):858-860
Objective To explore the signal transduction pathway of cyst fluid of Echinococcus granalosus in anti-parasite mechanisms through investigating the effect of cyst fluid on the expression of Toll-like receptor(TLR) in cells. Methods Changes of TLR4 and TGF-β1 expression of 7404 liver cells were detected by quantitative PCR. Results After treatment with increasing concentration cyst fluid the expres-sion of TLR4 was reduced. TGF-β1 expression of liver cells increased with the dose. TLR2 expressions in each group were very low. Conclusion Cyst fluid can increase the expression of TLR4, suggesting that the TLR4 signal transduction pathway involve anti-cyst fluid of Echinococcus granulosus. High concentrations of cyst fluid contribute to TGF-β1 expression which plays a role in immune evasion.
9.Effect of URG11 gene silencing on proliferation and invasion abilities of human prostate cancer cells
Bin PAN ; Zhihai DENG ; Youke WU ; Weibo LIANG ; Chuanrong ZENG ; Haiping LIU
Chinese Journal of Pathophysiology 2016;32(4):658-664
AIM:To investigate the expression of up-regulated gene 11 ( URG11 ) in prostate cancer cell line and the effect of URG11 siNRA on the proliferation and invasion of human prostate cancer LNCaP cells.METHODS:The mRNA and protein levels of URG11 in prostate cancer cell lines and normal prostate epithelial cell line were evaluated by real-time PCR and Western blot.LNCaP cells were transfected with designed siRNA using the liposome method.The prolif-eration, apoptosis, migration and invasion abilities of the LNCaP cells were evaluated by MTS assay, flow cytometry, wound-healing assay and Transwell assay.RESULTS:The expression of URG11 at mRNA and protein levels in the DU145, PC3, LNCaP cell lines was significantly higher than that in RWPE-1 cell line.Compared with the control group, the proliferation of LNCaP cells with URG11 siRNA was stagnant in G1/S phase and induced apoptosis.The proliferation of LNCaP cells at 0 h, 24 h, 48 h and 72 h was inhibited after URG11 expression was down-regulated (P<0.05).Transwell assay showed that migration (P<0.05) and invasion (P<0.05) were also inhibited.CONCLUSION:URG11 is highly expressed in prostate cancer.Silencing of URG11 significantly inhibits the proliferation and invasion and induces apoptosis of LNCaP cells.
10.Unilateral percutaneous vertebroplasty for osteoporotic vertebral compression fractures:less bone cement leakage and ideal recovery
Hong WU ; Yuan YUAN ; Lijin LIU ; Liang YAN ; Liwei XIONG ; Zhiyuan ZOU ; Zhihai MIN
Chinese Journal of Tissue Engineering Research 2015;(31):4960-4966
BACKGROUND:Percutaneous vertebroplasty and percutaneous kyphoplasty in the treatment of osteoporotic vertebral compression fracture have obtained good outcomes, because the traditional method is invalid, but there are a variety of choices in operation time, anesthesia, surgical approach and method, and each method has its advantages and disadvantages. OBJECTIVE:To investigate the effect and preponderance of the manual reduction combined with unilateral percutaneous vertebroplasty under general anesthesia in the treatment of osteoporotic vertebral compression fractures. METHODS:A total of 53 patients with single vertebral osteoporotic vertebral compression fractures, who were treated with percutaneous vertebroplasty, were retrospectively analyzed from July 2012 to December 2014. The new method group (32 cases) received manual reduction, underwent unilateral pedicle puncture and bone cement injection during unilateral percutaneous vertebroplasty under general anesthesia. The conventional method group (21 cases) received conventional percutaneous vertebroplasty. RESULTS AND CONCLUSION: There was an average of 6-month folow-up (3-14 months). Significant differences in visual analogue scale scores, vertebral compression ratio and kyphosis Cobb’s angle were detected in the new method and the conventional method groups at 3 days post surgery and during final folow-up compared with before surgery (P < 0.01). No significant difference in visual analogue scale scores was found between the two groups (P > 0.05). Compared with the conventional method group, postoperative vertebral compression ratio, kyphosis Cobb’s angle and bone cement leakage rate were significantly lower in the new method group (P < 0.01). Results verified that the new method combined with the advantages of percutaneous vertebroplasty and percutaneous kyphoplasty, the advantages of unilateral and bilateral puncture approach. The new method can correct kyphosis deformity, effectively recover the vertebral height and physiological curvature and the puncture is safe. Simultaneously, the leakage rate of bone cement is reduced, and the distribution of bone cement is ideal.