RESUMEN
In order to clean the endoscopic tube more effectively, a new double-headed endoscopic cleaning brush was used in this study. A total of 130 colonoscopies were selected from the Digestive Endoscopy Center of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine from June 2019 to August 2020. The colonoscopy cleaning sequence was marked with odd and even number. Colonoscopies marked with the odd number were assigned to the conventional group ( n=65) which received back and forth cleaning with the single-headed endoscope cleaning brush. Colonoscopies marked with the even number were assigned to the experimental group ( n=65) which received one-way cleaning with a nylon brush at the head and a dense non-woven brush at the tail. The cleaning methods for endoscopes were in accordance with Flexible Endoscopic Cleaning and Disinfection Technical Specification WS507—2016. ATP bioluminescence tests and bacterial quantitative cultures were applied to evaluate the cleaning effects of the two methods. The results showed that ATP relative light unit (RLU) decreased in both the experimental group and the conventional group [530.63 RLU (26-3 559 RLU) VS 270.87 RLU (20-1 415 RLU)] before and after cleaning the endoscope tube, showing significant difference between the two groups ( Z=-2.894, P<0.05). After scrubbing, the positive rate of bacterial culture on the brush head was 86.2% (56/65) for the double-headed non-woven brush head, 46.2% (30/65) for the double-headed nylon brush head and 32.3% (21/65) for the single-headed nylon brush head with significant differences among the three groups ( χ 2=41.046, P<0.05). The cleaning effect of the new double-headed endoscopic cleaning brush is better than that of the conventional single-headed endoscopic cleaning brush for soft endoscope cleaning. Non-woven brush is better than nylon brush in eliminating bacteria.
RESUMEN
Objective To analyze the prevalence of Clostridium difficile infection (CDI) in patients with inflammatory bowel disease (IBD) accompanied with diarrhea in order to analyze the possible risk factors and its impact on short-term prognosis.Methods From January 2013 to June 2015,a total of 169 IBD patients with diarrhea who visited Rui Jin Hospital,School of Medicine,Shanghai Jiaotong University were enrolled,and in the same period 184 non-IBD patients with diarrhea were enrolled as control group.Clinical data of IBD patients with diarrhea were collected.Clostridium difficile toxins A and B of stool specimens of all included patients were detected with enzyme immunoassay.T test,chi-square test and logistic regression analysis were performed for statistical analysis.Results Among 169 IBD patients with diarrhea,there were 137 adults and 32 children.The positive rate of CDI of IBD patients with diarrhea (9.5％,16/169) was higher than that of non-IBD patients with diarrhea (1.1％,2/184),and the difference was statistically significant (x2 =12.785,P ＜ 0.0l).The positive rate of CDI in adults and children with IBD accompanied with diarrhea were 7.3％ (10/137),and 18.8％ (6/32),respectively.Among 16 CDI-positive IBD patients with diarrhea,10 patients had hospitalization history before admission,and the rate of previous hospitalizations was higher than that of CDI-negative IBD patients with diarrhea (37.3％,57/153),and the difference was statistically significant (x2 =2.875,P =0.01,odds ratio (OR) =1.26,95％ confidence interval (CI) 0.78 to 2.03).Furthermore,among 16 CDI-positive IBD patients with diarrhea,14 patients had been treated with antibiotic drugs before,the antibiotic utilization rate was higher than that of CDI-negative IBD patients with diarrhea (34.6％,53/153),and the difference was statistically significant (x2 =14.778,P ＜ 0.01,OR =24.74,95％ CI 3.15 to 194.46).However,there was no statistically significant difference in the length of hospitalization,incidence of bowel surgery and usage of inflaximab within six months after Clostridium difficile detection between the CDI-positive group and CDInegative group of 1BD patients with diarrhea (all P ＞ 0.05).Conclusions The incidence of CD1 in IBD patients with diarrhea increases.The risk factors include history of previous hospitalization before admission and antibiotic usage.However there is no significant correlation between CDI and short-term prognosis of IBD.
RESUMEN
Objective To investigate the mechanism of TNF-related apoptosis-inducing ligand (TRAIL) promoting apoptosis of pancreatic cancer cells SW1990, Patu8988 and BxPC3. Methods Three kinds of pancreatic cancer cells SW1990, Patu8988 and BxPC3 were transfected with the pCA13 plasmid carrying TRAIL gene ( pCA13 TRAIL group) and the blank plasmid control ( pCA13 group) , respectively. The expression of TRAIL mRNA in transfected cells was detected by RT-PCR, and the expression of TRAIL protein was detected by Western blot. The apoptosis rate and expression of TRAIL receptor R1 and R2 were detected by flow cytometry. Apoptosis was detected by TUNEL and Hoechst double staining, and observed by electron microscopy. The expression of caspase-3 in transfected cells was detected by immunohistochemistry. Results SW1990, Patu8988 and BxPC3 cells can expresse TRAIL mRNA and protein within 24 h after transfection. The apoptotic rate at 24 h after transfection was (27. 30 ± 5. 14)％, (13. 52 ± 0. 95)％ and (31. 40 ± 8. 70)％,respectively, which was higher than that of pCA13 group [(10. 58 ± 1. 88)％,(8. 42 ± 0. 46)％ and (16.11 ±1.66)％], respectively. The expression rates of TRAIL-R1 were (61.37 ± 3.05)％,(42.10 ± 5. 11)％ and (36. 64 ± 4. 84)％, respectively, and the expression rates of TRAIL-R2 were (36. 20 ± 4. 83)％,(37. 26 ± 8. 46)％ and (24. 32 ± 3. 71)％, respectively,which were higher than those of pCA13 group except PATU8988 cells. Positivity rates of caspase-3 were ( 14. 64 ± 5. 35 )％, ( 9. 92 ± 5. 50 )％ and (16. 12 ± 6. 74)％, which were obviously higher than ( 3. 01 ± 1. 50 )％, ( 1. 75 ± 0. 50 )％ and ( 3. 79 ± 1. 58)％ in pCA13 group,and the differences were statistically significant(P<0. 05). Conclusions TRAIL could up-regulate the expression of TRAIL R1 and R2 in multiple pancreatic cancer cell lines in vitro, and thus promote cell apoptosis.
RESUMEN
OBJECTIVE:To observe therapeutic efficacy and safety of infliximab (IFX) in the treatment of Crohn's disease (CD),and to investigate its influential factors. METHODS:In retrospective study,a total of 120 CD patients received IFX treat-ment in our hospital during Jan.-Dec. 2015 were selected as research objects. All patients were given IFX for injection 5 mg/kg intra-venously at 0,2nd,6th week and later every 8 weeks continously. Therapeutic efficacy indexes(body weight,CRP,ESR,WBC, mucosal healing under endoscope),liver and renal indexes (ALT,AST,AKP,γ-GT,TB,DB,Cr),the occurrence time of ADR,organs/systems involved,clinical manifestations and outcomes were compared before and after treatment. Influential factors were analyzed. RESULTS:Compared with before medication,body weight of patients were increased significantly,while ESR were decreased significantly,with statistical significance (P<0.05). There was positively correlation between above two indexes (r=0.275,P=0.016). Mucosal healing was found in 78.57％ of patients by colonoscopy. There was statistical significance in the levels of AST,AKP and Cr before and after treatment(P<0.05),but those indexes were increased slightly. AKP exceeded 2 times the upper limit of normal value in only 1 case,but AST did not exceed 2 times the upper limit of normal limit,Cr did not exceed the upper limit of normal value. There was no statistical significance in CRP,WBC or other liver and renal function indexes before and af-ter treatment(P>0.05). There were 24 cases of ADR with incidence of 20.00％. Respiratory system,digestive system,skin mucous membrane,body defense and other organs/systems were involved,mainly manifestating as uper respiratory tract infetion,liver func-tion indexes order etc. Ten patients(41.67％)delayed the treatment,and 6 cases(25.00％)discontinued. Standard rate of ESR in pa-tients with regular/primary medication (78.21％) was higher than those with intermittent medication history (57.14％),but without statistical significance(P>0.05). Combined use of mesalazine had no significant effect on the increase of AST and AKP(P=0.002). CONCLUSIONS:IFX significantly increases body weight of CD patients,decreases ESR and improves mucosal healing. It shows def-inite therapeutic efficacy. It is suggested that CD patients should use IFX regularly. The overall safety of IFX is good,but it may cause transfusion reaction and transient increase of liver function index. So,the patient's condition should be closely monitored during the in-fusion.
RESUMEN
The prevalence of inflammatory bowel disease (IBD) in China is increasing year by year, however, the efficacy and safety of commonly used therapeutic methods are limited.Granulocyte and monocyte adsorptive apheresis (GMA) is one of the effective methods for treatment of IBD used abroad, however, there is still lacking of such research in China.Aims: To investigate the efficacy and safety of GMA in IBD patients.Methods: A retrospective study was conducted in 21 cases of IBD patients [13 cases with ulcerative colitis (UC) and 8 with Crohn's disease (CD)] who accepted GMA treatment from May 2013 to July 2014 at the Shanghai Rui Jin Hospital.All the cases were poor responders to 5-aminosalycylic acid (5-ASA) or steroid-refractory.The clinical data were collected, and the clinical activity index (CAI), endoscopic activity index (EAI), laboratory parameters including serum albumin (Alb), hemoglobin (Hb), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), leukocyte count and percentage of neutrophils, as well as the adverse effects before and two weeks after the end of GMA treatment were analyzed.Results: After GMA treatment, both CAI and EAI were decreased significantly in UC and CD groups as compared with those before treatment (P all 0.05).The treatment was well tolerated with no severe adverse effects.Conclusions: GMA is safe and effective for ameliorating clinical symptoms, attenuating intestinal mucosal injury and controlling active inflammation in IBD patient that has not responded to 5-ASA or steroid treatment.Prospective clinical studies with large samples are needed to confirm these findings.
RESUMEN
Objective To investigate the role of mucosal mast cells infiltration and degranulation with nerve growth factor (NGF)in development of visceral hypersensitivity in Sprague-Dawley (SD)rats. Methods The model of visceral hypersensitivity of irritable bowel syndrome (IBS)was established in 19 neonate SD rats with intestinal stimulation (rectalballon distention)on 8th,10th and 12th postnatal days. The other 19 neonate SD rats without colonic distention were assigned to the control group.After rats grew up (six to eight weeks old),the visceral sensitivity was tested by abdominal withdrawal reflex (AWR)in 10 rats of each group.Mast cell infiltration and degranulation were observed with toluidine blue staining in colon tissue slides.The NGF level of intestinal tissues was detected by enzyme-linked immunosorbent assay (ELISA)methods in the left nine rats of each group.The culture system of dorsal root ganglias (DRG)from the neonatal rats was set up.The changes of electrophysilogical characters of DRG stimulated with NGF (100 ng/mL)for four days were recorded with patch-clamp.Paired t test was performed for comparison between groups.Results The results of AWR indicated that neonatal colonic stimulation could significantly increase visceral sensitivity after growing up.Under 20,40 and 60 mmHg (1 mmHg=0.133 kPa)distention pressure,visceral sensitivity scores of visceral hypersensitivity rats and rats of control group were 1 .00±0.50 vs 1 .67 ±0.50,1 .89 ±0.31 vs 2.89 ±0.34 and 2.89 ±0.33 vs 3.89±0.33,the differences were statistically significant (t=-2.83,-6.00 and -6.00,all P <0.05 ). The results of master cells staining in tissue slides showed colonic master cells infiltration was obvious in rats with visceral hypersensitivity,and part of mast cells were degranulation.The result of ELISA demonstrated that NGF level of visceral hypersensitivity rats was significantly higher than that of control group ((11.07±3.06)pg/mg vs (2.38 ±1.88)pg/mg,t =-6.93,P <0.05).The results of electrophysilogical tests of primary cultured DRG indicated that compared with blank control growp,the action potential threshold of neuron in NGF 100 ng/mL group significantly decreased ((-18.0±2.1 )mV vs (-29.0 ± 2.5 )mV,t = 12.26,P <0.05)and discharge frequency increased ((5 .0± 1 .4 )/800 ms vs (12.0 ± 3.2)/800 ms,t=-8.40,P <0.05 ).Meanwhile,neuron voltage-gated K+ current density remarkably decreased,most were sustained delayed rectifier K+ current (I K )decreasing ((279.0 ±48.0)pA/pF vs (203.0±39.0)pA/pF,t=6.18,P <0.05).Conclusion Colonic stimulation in neonatal rats could cause intestinal master cells infiltration and degranulation,which induced changes of neuron electrophysilogical characters and resulted in visceral hypersensitivity after growing up.
RESUMEN
Objective To investigate the expression ot XAF1 protein in pancreauc cancer,and its relationship with clinicopathological parameters and prognosis.Methods The tissue microarray was composed of 89 pancreatic cancer and 21 normal pancreatic tissues,and immunohistochemistry was used to examine the expression of XAF1 protein.The correlation between XAF1 expression and clinicopathological parameters was analyzed.The 89 patients were followed,and the survival was shown in Kaplan Meier curve.Cox proportional hazards regression was used to identify risk factors for the survival of pancreatic cancer patients.Results The negative expression rate of XAF1 was 44.9％ (40/89),and it was 9.5％ (2/21) in normal tissue,and the difference between the two groups was statistically significant (P ＜0.05).The expression of XAF1 was negatively associated with tumor staging (P ＜ 0.05),but it was not associated with age,gender,lymph node metastasis,distant metastasis,pathological grading,surgical margins,vascular and neural invasion.Patients with negative expression of XAF1 had much shorter survival than patients with positive expression.Univariate analysis showed that negative expression of XAF1,pathological grading,lymph node metastasis,distant metastasis was associated with the survival (P ＜ 0.05),and the multivariate analysis indicated that negative expression of XAF1 was an independent prognostic factor for survival of pancreatic cancer.Conclusions XAF1 may be involved in the development and growth of pancreatic cancer,and it is related with patient's prognosis.
RESUMEN
Objective To investigate the circulating microRNAs carrier in pancreatic cancer.Methods Pancreatic cancer cell lines SW1990 and BxPC3 were routinely cultured and serum of 6 patients with pancreatic cancer and 6 healthy subjects (control group) were collected.Serum and pancreatic cancer cell line supernatant microvesicles (MV) were obtained by gradient centrifugation.The expression of Ago2,CD63 was detected by Western blotting,the expression of microRNA in microvesicle section and microvesicle-free section in serum was detected by using quantitative PCR method.Results The supernatant MV of pancreatic cancer cell lines expressed Ago2,CD63 protein,and these MV carried different microRNAs in different cell lines.In the serum of pancreatic cancer and control group,miR-20a,miR-21,miR-24,miR-25,miR-191,miR-483-5p were detected,but the quantity was relatively higher in MV section,and the expression of microRNAs in pancreatic cancer's MV was inconsistent with that of control group.The expression of miR-20a,miR-24,miR-191 in pancreatic cancer group was (2.93 ± 0.29),(2.73 ± 0.46),(2.39 ± 0.51) times as much as those in control group,and the difference between the two groups was statistically significant (F =75.97,25.80,12.94,P ＜ 0.05 or ＜ 0.01).Conclusions The main circulating microRNAs carrier in pancreatic cancer is microvesicle.
RESUMEN
Objective To look for and confirm the downstream regulated gene of microRNA (miRNA)-148a in pancreatic cancer cell line.Methods The target gene regulated by miRNA-148a was predicted through bio-informatics analysis.The plasmid containing desired gene and with luciferase 3'-untranslated region (3'-UTR) reporter was constructed.Pancreatic cancer cells BXPC-3 were transfected with analogs and inhibitors of miRNA-148a by liposomes.The activity of luciferase was measured to determine whether miRNA-148a directly connected with desired gene.The expression level of miRNA-148a was changed in BXPC-3 cells,and the changes of target gene v-verb-b2 erythroblastic leukemia viral oncogene homolog 3 (awian) (ErbB3) expression were detected by Western blot at protein level.The data were analyzed by one way ANOVA.Results There was a conservative binding site of ErbB3 with miRNA-148a detected by bio-informatics analysis,miRNA-148a directly combined with ErbB3 and the activity of luciferase decreased to (25.00+47.00) ％ of the negative control (F=4.66,P＜ 0.01).After miRNA-148a overexpression,the gray value of ErbB3 expression in BXPC-3 cells decreased to (26.16±4.69)％ of control group (F=6.563,P＜0.05).Conclusion miRNA-148a directly targeted and regulated the expression of ErbB3 in pancreatic cancer cell line BXPC-3.
RESUMEN
Objective To investigate the diagnostic value of UL-16 binding protein 2 (ULBP-2,macrophage inhibitory cytokine-1 (MIC-1) for pancreatic cancer.Methods The serum samples of 152pancreatic cancer patients,20 precursors of pancreatic cancer,91 chronic pancreatitis patients and 96 age/sexmatched healthy persons were collected.The serum ULBP-2 and MIC-1 levels were determined by using the ELISA kit and were compared with level of CA19-9.A receiver operating characteristic (ROC) curve was constructed to evaluate their diagnostic values for pancreatic cancer.Results The serum levels of ULBP-2 in patients with pancreatic cancer,precursors of pancreatic cancer,chronic pancreatitis and healthy persons were (219.9 ± 182.5),(62.6 ± 11.4),(68.4 ± 36.8),(76.5 ± 40.9) μg/L,the corresponding values of MIC 1 were (3521.3±3903.4),(973.6±589.0),(959.6±879.0),(427.6±317.0) μg/L,while the corresponding values of CA19-9 were (1448.8 ± 3707.0),(12.0 ± 9.3),(38.2 ± 139.0),(7.7 ± 5.0)kU/L.The parameters in pancreatic cancer patients were significantly higher than those in control group (x2 =40.628,71.662,45.505,15.827,36.433,63.494,26.264,73.427,49.088,P ＜ 0.01).The area under ROC curves(AUC) of ULBP-2,MIC-1,CA19-9 were 0.909,0.864,0.818,and ULBP-2 was superior to CA19-9 and MIC-1,however the combined measurement of three markers produced the highest diagnostic yield(AUC =0.982).For early stage pancreatic diseases (precursors to pancreatic cancer and IA stage pancreatic cancer),AUC of ULBP-2,MIC-1,CA19-9 were 0.506,0.837,0.684,MIC-1 was superior to ULBP-2 and CA19-9,however the combined measurement of MIC-1 and CA19-9 produced the highest diagnostic yield(AUC =0.897).Conclusions Serum ULBP-2,MIC-1 levels are significantly elevated in pancreatic cancer patients.The combined measurement of ULBP-2,MIC-1 and CA 19-9 can increase the diagnostic yield for pancreatic cancer.
RESUMEN
ObjectiveTo investigate the expression of protein inhibitor of activated STAT-1 ( PIAS1 )in rat with acute pancreatitis (AP) and study its prediction value for severity and prognosis.MethodsSD rats were randomly divided into control,acute edematous pancreatitis (AEP),and acute necrotizing pancreatitis (ANP) groups with 15 rats in each group.The rats were sacrificed at 0.5,6,16,24,48,72 h postoperatively.Serum CRP and amylase levels were determined.Water content of pancreas was measured.The pancreatic tissue was routinely harvested and underwent pathologic examination and was scored.The PIAS1protein expression was measured by Western blot and immunohistochemistry method.The survival rates at 72h were recorded and the risk analysis of multiple factors was investigated by Cóx regression method.ResultsThe pathologic score,water content of pancreas,serum CRP and amylase levels at 16h in ANP group were (7.70 ±2.55),(2.91 ±0.57)％,(0.36 ±0.05)mg/L,(3141 ±625)U/L,which were significandy higher than those in AEP group [(2.60 ±1.04),(2.04 ±0.47)％,(0.24 ±0.05)mg/L,(1984 ± 637)U/L)],and also significantly higher than those in control group [ (0.80 ±0.42),( 1.21 ±0.27)％,(0.14 ±0.03)mg/L,(978 ± 353) U/L,(P ＜ 0.05or 0.01 ) ].The survival time of ANP rats was [ (26.36 ± 3.41 ) h,95％ CI:19.67～33.06 h],which were significantly shorter than that in AEP group [ (57.26 ±4.16)h,95％ CI:49.11 ～ 65.42) ],and also significantly shorter than that in control group [ (71.33 ±0.54) h,95％ CI:70.26 ～71.40,P ＜0.01 ].The expression of PIAS1 at 16h in ANP group was significantly lower than those in AEP group and control group ( 0.10 ± 0.01 vs.0.80 ± 0.07,0.87 ± 0.05,P ＜ 0.01 ).The Cox analysis suggested that PIAS1 was an independent prognosis factor for the survival time of AP rats.ConclusionsPIAS1 is lowly to moderately expressed in ANP,and is negatively correlated with AP severity,and may be an independent risk factor for the prediction of severity and prognosis of AP.
RESUMEN
Objective To investigate the expression of actin-associated protein Transgelin in pancreatic cancer with or without diabetes and its effects on migration and invasion in SW1990 cell line. Methods The expression of Transgelin in 92 pancreatic cancer tissue specimens (45 cases accompanied with diabetes) and adjacent tumor-free tissue specimens (over 5cm from the edge of the tumor) was detected by immunohistochemistry, and their association with clinical pathological characteristics were also analyzed. Transgelin siRNA was designed and transfected into pancreatic cancer cell line SW1990. The changes of migration and invasion before and after transfection were observed through Transwell test.Results The positive percentage of Transgelin expression in pancreatic cancer was 68.5 ％ (63/92), which was significantly higher than that of adjacent tumor-free tissues[33.7％ ( 31/92), P＜ 0.05]. The positive percentage of Transgelin expression in pancreatic cancer accompanied with diabetes was 84.4％ (38/45), which was significantly higher than that without diabetes[53.2％ (25/47), P＜0.05]. The expression of Transgelin in pancreatic cancer tissues was associated with lymph nodes metastasis and TNM staging (both P＜0.05), but not related with gender, age, site, differentiation and portal vein or nerve invasion (P＞0.05). After Transgelin was interfered for 48 hours, the migration ability was significantly lower (migration cell number 49.2 ±9.5 cells) than negative control group (61.9±7.5 cells) and blank group (65.3±10.6 cells) (both P＜0.05), and the invasion of SW 1990 cells (48.0 ± 8.6 cells) also significantly lower than negative control group (63.5±11.4 cells) and blank group (67.5±9.6 cells) (both P＜0. 05). Conclusion Transgelin may involve in the metastasis of pancreatic cancer accompanied with diabetes through promoting pancreatic cancer cell migration and invasion.
RESUMEN
Objective To determine the serum KL-6 level in patients with pancreatic carcinoma and investigate its diagnostic value. Methods The data of 53 pancreatic carcinoma (PC) patients; 68 chronic pancreatitis (CP) patients and 51 patients with high risks of pancreatic cancer (HR) with complete follow-up data were studied, and 50 healthy volunteers were used as controls. ELISA was used to measure the serum levels of KL-6, MUC4, and CA50. Radioimmunoassay methods were used to measure the serum CA19-9 levels. The sensitivity and specificity of KL-6 for the diagnosis of PC were determined, and the relationship between its levels and clinicopathologic parameters, patients' outcome were analyzed. Results The serum KL-6 levels were (753±548), (135±93), (105±55) and (99±50) U/ml in PC group, CP group, HR group and control group, and the value in PC group was significantly higher than those in other 3 groups (P 232 U/ml as the cut-off point, the sensitivity and specificity of KL-6 for the diagnosis of PC was 96% and 94%. Using >244 U/ml as the cut-off point, the sensitivity and specificity of KL-6 for the diagnosis of PC was 97% and 91%. The clinical value of KL-6 was higher than those of CA19-9, CASO and MUC4. The serum level of KL-6 was not associated with clinicopathologic parameters. The median survival of patients with serum level of KL-6 =300 U/ml was (9.3 ±1.2) months, which was significantly longer than that in patients with serum level of KL-6 >300 U/ml [ (4.6 ±0.7) months, P =0.006]. Conclusions KL-6 might be a useful serological marker of pancreatic cancer, and it may play a role in the differential diagnosis between pancreatic cancer and chronic pancreatitis.
RESUMEN
Objective To screen the difference of gene expression in dorsal root ganglia (DRG)of inflammatory visceral hypersensitivity rats and to explore the role of voltage gated calcium channel (VGCC) in inflammatory visceral hypersensitivity. Methods Total 180 male Sprague-Dawley rats were in this study,the weight varied from 200 gram to 300 gram. 2,4,6-trinitrobenzenesulfonic acid (TNBS) model group was maken by 2. 0% TNBS slowly injection,the dosage was 100mg per kilogram. The normal control group was only injected with same volume of 0. 9% sodium chloride solution. After the model had been maken for four days,gene expression profiles of L6-S2 DRGs were tested by rat cDNA microarray chips. And the result was verified by RT-PCR and Western blot. The changes of intracellular Ca2+ and the voltage gated calcium currents were recorded by patch-clamp.The special Ca2+ channel blockers were given by intrathecal injection,and then the changes of visceral sensitivity were observed. The visceral sensitivity was measured by abdominal withdrawal reflex (AWR). Results There were significant changes of 172 genes expression in L6-S2 DRGs of TNBS model rats,which included Ca2+ channel,membrane receptor and intracellular second messenger. Of those,L-type Ca2+ channel (Cav1. 2) and R-type Ca2+ channel (Cav2. 3) were significantly up-regulated. The results of gene microarray chips were further confirmed by RT-PCR and Western blot.The intracellular Ca2+ testing indicated that there was no statistical significant of resting intracellular Ca2+ in colonic special sensory neuron between TNBS group and normal control group (P＞0. 05);while the evoked transients [Ca2+] significantly increased compared with normal control group (P＜0. 05). The whole cell patch clamp recording showed that the L-type and R-type calcium current were significantly increased in colonic primary sensory neurons of TNBS group compared with normal control group (P＜0. 05). The inflammatory visceral hypersensitivity was significantly reduced by intrathecal injection of nimodipine and SNX-482 (P＜0. 05). Conclusion The up-regulation of Cav1. 2and Cav2. 3 play an important role in inflammatory visceral hyperalgesia,which may be the possible potential therapeutic targets for visceral inflammatory hyperalgesia.
RESUMEN
Objective To investigate the relationship between the activity of Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway in cerulein-induced pancreatic acinar cell line AR42J. Methods The in vivo model of AP was induced by cerulean treated pancreatic acinar cell line AR42J, then RPM and AG490 were given for intervention. Western blot was used to determine theexpressions of JAK1 and phosphorylation JAK1 ( P JAK1 ) , STAT1, PSTAT1 and TNFα, IL-1β, IL-6. The expressions of IL-6, IL-1 β, and TNFα mRNA were measured by RT-PCR. Survival rate of cells was evaluated by trypan blue stain. Results The relative expressions of JAK1, P JAK1, STAT1, P STAT1 and TNF-o, IL-1β, IL 6 without cerulean treatment were 0.09 ±0.04,0.14 ±0.08,0.21 ±0.09,0.12 ±0.12,0.10 ±0.02,0.08 ± 0.03,0.02 ± 0.02. After cerulean treatment, the expressions of abovementioned protein increased in a time-dependant manner, the expressions at 24h were 0.53 ± 0.09,0.53 ± 0.13,0.56 ± 0.09,0.55 ± 0.10,0.25 ± 0.04,0.25 ±0.09,0.27 ±0.07, which were significantly higher than those in the control group (P ＜0.05). 2 4 h after RPM and AG 4 9 0 inhibition, the expressions of TNF-α, IL-1 β, IL-6 proteins significantly decreased to 0.17 ± 0.03and 0.17 ± 0.01,0.15 ± 0.05 and 0.14 ± 0.07,0.19 ± 0.04 and 0.19 ± 0.05; their expressions of mRNA significantly decreased ( P ＜ 0.05 ). The cell survival rates in RPM and AG490 treatment group were (72.4 ± 11.2) %, (69.7 ± 9.8 ) %, and in cerulein-stimulated cells (42.2 ± 12.3 ) % ( P ＜ 0.05 ).Conclusions The JAK1/STAT1 signaling pathway was involved in pancreatic inflammatory response with cerulein stimulation. Early treatment with inhibitors to the JAK1/STAT1 signaling pathway might control the inflammatory response in acute pancreatitis.
RESUMEN
Objective To provide an objective basis for differential diagnosis of pancreatic diseases through quantitative analysis of the different features of contrast-enhanced endoscopic ultrasonography (CE-EUS). Methods A total of 32 patients with suspected or confirmed pancreatic neoplasms or chronic pancreatitis and 19 patients who underwent EUS due to other digestive problems other than pancreatic disease were enrolled. Features of blood perfusion of the regions of interest during CE-EUS were analyzed quantitatively. The findings were compared with cytological and/or histopathological results of EUS-FNA and/or surgery.Results Quantitative analysis of CE-EUS showed peak intensity (PI) value of 19 normal pancreas was 0.648 ±0. 174, which was statistically different from that of pancreatic cancer and pancreatic cystic lesions. Based on ROC, the cutoff of differential diagnosis was 0. 505, and the sensitivity and specificity were 100. 0% and 84. 2%, respectively. PI value of 6 chronic pancreatitis was the highest (0. 772 ±0. 106). In pancreatic neoplams, PI values of pancreatic carcinoma, pancreatic cyst and pancreatic endocrine tumors were significantly different. Based on a cutoff of 0. 195, the sensitivity and specificity of differentiation of pancreatic carcinoma and pancreatic cyst were 85.7% and 87.5%, respectively. PI value of 14 pancreatic carcinoma and that of 4 pancreatic endocrine tumors were 0. 321 ± 0. 119 and 0. 763 ± 0. 115, respectively. Through the comparison between the AT and TTP of the focal lesions and surrounding pancreatic parenchyma, 78.6% pancreatic carcinoma showed slow falling-in and rapid wash-out and all the endocrine tumors showed rapid falling-in and rapid wash-out. The PI value of 8 patients with pancreatic cyst was 0. 181 ±0. 036, with no enhanced blood flow in the cyst. The TIC was a straight line. Conclusion CE-EUS with quantitative analysis is a promising method that can be a more objective basis in the differential diagnosis of pancreatic diseases.
RESUMEN
Objective To analyze the relationship between blood glucose level, blood pressure and weight with pancreatic cancer genesis. Then to explore the metabolism associated risk factors in pancreatic cancer genesis. Methods Form December 2002 to September 2009 in Ruijin Hospital, 548 pancreatic cancers with pathology diagnosis after pancreatectomy were collected for the study with retrospective analysis method. The association of pancreatic cancer with blood glucose level, blood pressure, weight and other metabolic factors were analyzed. Results With principal component analysis, it suggested that there were strong correlation between blood glucose level, blood pressure and weight index (BMI) increasing with pancreatic cancer. The contribution rates were 3. 614%,25. 236%, 15. 418% and 12. 918%, respectively. Single factor analysis indicated that the association between pancreatic cancers and new onset diabetes mellitus (duration≤ 2 years) was stronger than that of long-term diabetes mellitus. The occurrence rate of pancreatic cancer in patients with long-term diabetes whose blood glucose level was not well controlled recently while well controlled previously (44.6 % ) was significant hister than that in patients without diabetes (5. 6% , P＜0.05). The fasting blood glucose level of these PC patients ( 13.87± 3. 49 mmol/L) was significantly higher than new onset and other long-term diabetes patients, the comparative risk was 13.46 (95% CI 4. 560,39. 731). BMI increasing was a risk factor of pancreatic cancer, but there was no significant statistical difference between risk degree and BMI increasing level. All above metabolic diseases were risk factors of pancreatic cancer, but for pathology, location and stage of pancreatic cancer there was no statistical difference in theses factors. Conclusion This study suggested diabetes, BMI increasing and hypertension were high risk factors of pancreatic cancer genesis. New onset and long-term diabetes patients whose blood glucose not controlled well recently should be watched carefully for pancreatic cancer. Early treatment and intensive follow-up of metabolic disease might be helpful to early diagnosis and prognosis of pancreatic cancer.
RESUMEN
Objective To investigate the effect of protein inhibitor of activated signal transducer and activator of transcription 1 ( PIAS1 ) gene silencing on the inflammatory response of rat pancreatic acinar cell lines AR42J with cerulein stimulation, to study its role in the pathogenesis of acute pancreatitis.Methods The siRNA targeting PIASI was designed, synthesized, transfected into AR42J cells by lipofectmine 2000.24 h later, cerulean was added and cultured for another 24 h.Subsequent AR42J cells with cerulein stimulation were divided into 4 groups: cerulein, liposome, negative-siRNA and PIAS1-siRNA groups.In addition, a group with PBS was as control group.The activity of p38 mitogen- activated protein kinase (p38MAPK) was detected by western blotting.TNF-α, IL-1β, IL-6, matrix metalloproteinase (MMP) 9 expression were analyzed by RT-PCR and western blotting, respectively.Results The expression of p38MAPK in PIAS1-siRNA, negative-siRNA, liposome, cerulein,and control group was 1.93 ±0.11, 1.22 ±0.10, 1.30 ±0.17,1.32 ± 0.21, 0.12 ± 0.02;while the expression of phosphorylated p38MAPK was 2.10 ± 0.25, 1.36 ± 0.20,1.26 ±0.15, 1.23 ±0.25, 0.58 ±0.48, the expression in PIAS1-siRNA group was significantly increased when compared with other groups (P＜0.05).The levels of TNF-α, IL-1β, IL-6, MMP-9 mRNA were 1.66 ±0.15,1.66 ± 0.15,1.90 ±0.01, 1.56 ±0.20 in PIAS1-siRNA group, while the expression of protein was 2.06 ±0.37,2.20 ±0.34, 1.80 ±0.10, 1.17 ±0.05, which was markedly higher than those in other group (P ＜0.05).Conclusions PIAS1 gene silencing could enhance p38MAPK activity, and improve inflammatory mediator expression in pancreatic acinar cells with cerulein stimulation.
RESUMEN
Objective To investigate the role of connexin 43(Cx43)in the apoptosis of pancreatic cancer cell line BxPC and its possible mechanism.Methods pcDNA-Cx43,pcDNA-Cx43N,pcDNA3.0,siRNA-Cx43 and siRNA-NC were transfected into BxPC3 cells via liposome method.Cx43 protein and Cytochrome C(Cyt C)concentration was determined by Western blot,and the apoptosis was analyzed by Annexin V/PI binding assay.The mitechondria apoptosis pathway involved in Cx43 associated apoptosis was examined which contains the depolarization of mitechondrial membrane potential (MMP);fluorospectrophotometer was used to measure the activities of caspase-3 and caspase-9. Gap junction intercellular communication(GJIC) was determined by dye-transfer method.Results Cx43 protein expression increased after BxPC3 transfeetion,apoptosis rate increased from(6.35±0.43)%in empty vector transfection group to(14.29±1.24)%;after H202 treatment,apoptosis rate increased from(20.34±2.47)%to(31.27±2.56)%(P<0.05).Meanwhile,mitochondrial membrane potential was decreased,Cyt C was increasingly released from mitochondria,caspases activities were increased;after siRNA43 interference,apoptosis rate decreased from(7.42±0.47)% to(5.19±1.37)%,after H_2O_2 treatment,apoptosis rate decreased from (19.43±1.71)%to(11.67±1.97)%(P<0.05).Decreased mitochondrial membrane potential and Cyt C release were observed,caspases activities were decreased.GJIC of pcDNA-Cx 43 transfection group increased from 14.52±0.57 to 23.05±3.84.and it increased from 1.70 ±0.24 to 3.84 ±0.45 in the presence of β-GA(P<0.05).But the apoptosis rate was not significantly different.Conclusions Cx43 could promote BxPC3 apoptosis via mitochondrial apoptotic signal pathway,and the possible mechanism included signal pathway other than GJIC.
RESUMEN
Objective To investigate whether expression of XAF1 mediated by edenovirus vector AdS/F35 could induce apoptosis in pancreatic cancer cell line BxPC-3 and its possible mechanisms.Methods Preconstructed recombinant Ad5/F35-XAF1 virus and negative control Ad5/F35-Null was tranfected into BxPC3:the expression of XAF1 mRNA and protein before and after tranfection was,analyzed by semi-quantitative RT-PCR and Western blot.The expressions of proteins including Caspase-3,PARP,Caspase-8 and bcl-2 were detected by Western blots.Cell apoptosis was assessed by Annexin-v/PI and TUNEL staining.Results After Ad5/F35-XAF1 tranfection,XAF1 mRNA and protein expression significantly increased,and the difference was statistically significant when compared with control group and Ad5/F35-Null group(P＜0.05).The apoptosis rate was(19.90±3.09)%and(9.29±2.13)%,which was significantly different(P＜0.01)from those in the Ad5/F35-Null group[(6.72±0.7)6%,(2.73±0.51)%]or in the control group[(7.22±1.53)%,(1.56±0.47)%].The expression of Caspase-3,PARP and Caspase-8 significantly increased,but the expression of bcl-2 decreased.Conclusions XAF1 plays a major role in the apoptosis of pancreatic cancer that acts thriugh the activation of death receptor pathway and mitochondrial pathway of apoptosis.