利用胰酶粉提取胰激肽释放酶静态条件的初探

研究利用胰酶粉提取胰激肽释放酶的静态提取工艺。分别考察了浸提pH值、平衡液pH值、吸附过程中的NaCl添加量、洗脱过程中pH值、NaCl添加量以及醋酸钠正交试验对利用胰酶粉静态提取胰激肽释放酶的影响。利用胰酶粉静态提取胰激肽释放酶的最佳条件为:浸提pH值为6.0,平衡液pH值为6.5,吸附过程中添加0.02mol/LNaCl,洗脱优化条件控制在pH值为6.5,NaCl0.35mol/L以及不添加醋酸钠,提取率达到76%以上,该静态条件的考察为胰激肽释放酶的进一步分离纯化提供了理论依据。

胰酶的制法

胰酶是由胰脏提制的酶制剂。主要含有胰蛋白酶、胰脂肪酶、胰凝乳蛋白酶等。工业用胰酶制剂,在皮革工业上用于鞣革、脱毛、脱脂;在纺织工业上用于脱浆等等。 (一) 主要原料及加工材料: (1) 猪、牛、羊的白胰及鸡、鸭、鹅的胰脏。

胶体磨在生化制品中的应用

近年来,我国在引进并消化吸收国外胶体磨技术基础上,廊坊通用机械厂开发研制出了超微研磨、乳化设备——双一牌JM系列胶体磨。并在生化制药生产中进行推广应用,大大提高了生化药品胰酶粉的收率和质量,并获得很高的经济效益和社会效益。加工原理JM系列胶体磨是超微粒综合加工的理想设备,可代替均质机、球磨机。

Drainage fluid and serum amylase levels accurately predict development of postoperative pancreatic fistula

AIM To investigate potential biomarkers for predicting postoperative pancreatic fistula( POPF) after pancreaticoduodenectomy(PD).METHODS We prospectively recruited 83 patients to this study. All patients underwent PD(Child's procedure) at the Division of Hepatobiliary and Pancreas Surgery at the First Bethune Hospital of Jilin University between June 2011 and April 2015. Data pertaining to demographic variables, clinical characteristics, texture of pancreas, surgical approach, histopathological results, white blood cell count, amylase and choline levels in the serum, pancreatic/gastric drainage fluid, and choline and amylase levels in abdominal drainage fluid were included in the analysis. Potential correlations between these parameters and postoperative complications such as, POPF, acute pancreatitis, hemorrhage, delayed gastric emptying, and biliary fistula, were assessed. RESULTS Twenty-eight out of the 83(33.7%) patients developed POPF. The severity of POPF was classified as Grade A in 8(28%) patients, grade B in 16(58%), and grade C in4(14%), according to the pancreatic fistula criteria. On univariate and multivariate logistic regression analyses, higher amylase level in the abdominal drainage fluid on postoperative day(POD)1 and higher serum amylase levels on POD4 showed a significant correlation with POPF(P < 0.05). On receiver operating characteristic curve analysis, amylase cut-off level of 2365.5 U/L in the abdominal drainage fluid was associated with a 78.6% sensitivity and 80% specificity [area under the curve(AUC): 0.844; P = 0.009]. A cut-off serum amylase level of 44.2 U/L was associated with a 78.6% sensitivity and 70.9% specificity(AUC: 0.784; P = 0.05).CONCLUSION Amylase level in the abdominal drainage fluid on POD1 and serum amylase level on POD4 represent novel biomarkers associated with POPF development.

Preventing pancreatic fistula after distal pancreatectomy: An invagination method

Following an increase in the use of the GIA stapler for treating a pancreatic stump, more techniques to prevent postoperative pancreatic juice leakage have been required. We describe one successful case using our new technique of invaginating the cut end of the pancreas into the stomach to prevent a pancrea-tic fistula(PF) from occurring. A 50-year-old woman with pancreatic cancer in the tail of the pancreas underwent distal pancreatectomy, causing a grade A PF. We resected the distal pancreas without additional reinforcement to invaginate the stump into the gastric posterior wall with single layer anastomosis using a 3-0 absorbable suture. The drain tubes were removed on the third postoperative day. Although a grade A PF was noted, the patient was discharged on foot on the eleventh postoperative day. Our technique may be a suitable method for patients with a pancreatic body and tail tumor.

N-acetylcysteine does not prevent post-endoscopic retrograde cholangiopancreatography hyperamylasemia and acute pancreatitis

AIM： Acute pancreatitis （AP） is the most common and often severe complication of endoscopic retrograde cholangiopancreatography （ERCP）. The early step in the pathogenesis of acute pancreatitis is probably the capillary endothelial injury mediated by oxygen-derived free radicals. N-acetylcysteine - a free radical scavenger may be potentially effective in preventing post-ERCP acute pancreatitis and it is also known that N-acetylcysteine （ACC） can reduce the severity of disease in experimental model of AP. METHODS： One hundred and six patients were randomly allocated to two groups. Fifty-five patients were given N-acetylcysteine （two 600 mg doses orally 24 and 12 h before ERCP and 600 mg was given iv, twice a day for two days after the ERCP）. The control group consisted of 51 patients who were given iv. isotonic saline twice a day for two days after the ERCP. Serum and urine amylase activities were measured before ERCP and 8 and 24 h after the procedure. The primary outcome parameter was post-ERCP acute pancreatitis and the secondary outcome parameters were differences between groups in serum and urine amylase activity. RESULTS： There were no significant differences in the rate of post-ERCP pancreatitis between two groups （10 patients overall, 4 in the ACC group and 6 in the controlgroup）. There were also no significant differences in baseline and post-ERCP serum and urine amylase activity between ACC group and control group. CONCLUSION： N-acetylcysteine fails to demonstrate any significant preventive effect on post-ERCP pancreatitis, as well as on serum and urine amylase activity.

Perforated appendicitis masquerading as acute pancreatitis in a morbidly obese patient

Diagnosis and treatment of common conditions in morbidly obese patients still pose a challenge to physicians and surgeons.Sometimes too much reliance is put on investigations that can lead to a misdiagnosis.This case demonstrates an obese woman admitted under the medical team with a presumed diagnosis of pneumonia, who was later found to have an acute abdomen and raised amylase,which led to an assumed diagnosis of pancreatitis.She died within 24 h of admission and post mortem confirmed the cause of death as systemic sepsis due to perforated appendicitis,with no evidence of pancreatitis.Significantly elevated serum amylase level may occur in non-pancreatitic acute abdomen.

Breviscapine attenuates acute pancreatitis by inhibiting expression of PKCα and NF-κB in pancreas

AIM:To study the effect of breviscapine (Bre) on activity of protein kinase Cα (PKCα) and nuclear factor (NF)-κB in pancreas,and the mechanism of Bre attenuating acute pancreatitis (AP). METHODS:One hundred and eight rats were randomly divided into acute necrotizing pancreatitis (ANP) group,Bre group (ANP + Bre group) and sham operation (SO) group,36 rats in each group. ANP model was induced by a retrograde injection of 4% sodium deoxycholate into the bilio-pancreatic duct. Fifteen minutes after the ANP model was induced,the rats in Bre group were intraperitoneally injected with Bre (0.4 mg/100 g body weight or 0.1 mL/100 g body weight). Survival time and mortality of rats were calculated. Serum amylase and malondialdehyde levels were measured,volume of ascites was recorded and morphology of pancreas and lung was evaluated at 1,5 and 10 h,after the ANP model was induced,respectively. Expressions of PKCα and subunit p65 of NF-κB in pancreas were detected by immunohistochemistry and Western blotting. RESULTS:The life span of rats was longer and the mortality was lower in Bre group than in ANP group 13.51 ± 5.46 vs 25.36 ± 8.11 (P < 0.05). The amylase and MDA levels as well as the volume of ascites were lower and the pathological changes in pancreas and lung were less in Bre group than ANP group (P < 0.05),indicating that the pancreatitis is less severe in Bre group than ANP group. The activation of PKCα and NF-κB p65 in pancreas was induced rapidly and reached their peak at 1 h or 5 h after ANP,but their activity in Bre group was significantly inhibited. CONCLUSION:Bre exerts its therapeutic effect on AP by inhibiting the activation of PKCα and NF-κB p65 in pancreas.

Relationship between post-ERCP pancreatitis and the change of serum amylase level after the procedure

AIM: To clarify the relationship between the change of serum amylase level and post-ERCP pancreatitis. METHODS: Between January 1999 and December 2002, 1291 ERCP-related procedures were performed. Serum amylase concentrations were measured before the procedure and 3, 6, and 24 h afterward. The frequency and severity of post-ERCP pancreatitis and the relationship between these phenomena and the change in amylase level were estimated. RESULTS: Post-ERCP pancreatitis occurred in 47 patients (3.6%). Pancreatitis occurred in 1% of patients with normal amylase levels 3 h after ERCP, and in 1%, 5%, 20%, 31% and 39% of patients with amylase levels elevated 1-2 times, 2-3 times, 3-5 times, 5-10 times and over 10 times the upper normal limit at 3 h after ERCP, respectively (level < 2 times vs ≥ 2 times, P < 0.001). Of the 143 patients with levels higher than the normal limit at 3 h after ERCP followed by elevation at 6 h, pancreatitis occurred in 26%. In contrast, pancreatitis occurred in 9% of 45 patients with a level higher than two times the normal limit at 3 h after ERCP followed by a decrease at 6 h (26% vs 9%, P < 0.05). CONCLUSION: Post-ERCP pancreatitis is frequently associated with an increase in serum amylase level greater than twice the normal limit at 3 h after ERCP with an elevation at 6 h. A decrease in amylase level at 6 h after ERCP suggests the unlikelihood of development of post-ERCP pancreatitis.

Acute pancreatitis and amiodarone:A case report

Amiodarone, a class Ⅲ antiarrhythmic drug, is one of the most effective drugs used in the treatment of ventricular and paroxysmal supraventricular tachyarrhythmia. Adverse effects of amiodarone including pulmonary toxicity, hepatotoxicity, aggravation of arrhythmia, and thyroid diseases are well understood. A 66-year old woman with acute pancreatitis was admitted to our hospital with the complaint of epigastralgia radiating to both flanks for two months. Her symptoms and elevation of pancreatic enzymes did not respond to conventional medical treatment of pancreatitis for 18 d. No known causal factors for pancreatitis such as biliary tract stone, hypertriglyceridemia and alcohol consumption could be identified. Under the suspicion of amiodarone-induced acute pancreatitis, amiodarone was substituted by propafenone. Her symptoms soon alleviated and serum lipase level declined. Three months after hospital discharge, the abdominal pain did not recur. Amiodarone was approved to treat recurrent ventricular fibrillation or sustained ventricular tachyarrhythmia that has been resistant to other medications since 1986. Pancreatitis is a very rare adverse effect associated with the use of amiodarone, and only four cases of amiodarone-induced pancreatitis have been reported in literature. We report a patient who developed acute pancreatitis during amiodarone therapy.

Types of voltage-dependent calcium channels involved in high potassium depolarization-induced amylase secretion in the exocrine pancreatic tumour cell line AR4-2J

In the perifused fura-2 loaded exocrine pancreatic acinar cell line AR4-2J pulses of high potassium induced repetitive increases in intracellular calcium. Attached cells when stimulated with high potassium secreted large amount of amylase. High potassium-induced secretion was dependent both on the concentration of potassium and duration of stimulation. High potassium induced increases in intracellular calcium were inhibited by voltage-dependent calcium channel antagonists with an order of potency as follows: nifedipine > ω-agatoxin IVA > ω-conotoxin GVIA. In contrast, the L-type calcium channel antagonist nifedipine almost completely inhibited potassium-induced amylase secretion, whereas the N-type channel antagonist ω-conotoxin GVIA was without effect. The P-type channel antagonist ω-agatoxin IVA had a small inhibitory effect, but this inhibition was not significant at the level of amylase secretion. In conclusion, the AR4-2J cell line possesses different voltage-dependent calcium channels (L, P,N) with the L-type predominantly involved in depolarization induced amylase secretion.

Synergetic Effect of Yihuo Qingyi Decoction (益活凊胰汤) and Recombinant Staphylokinase in Treatment of Severe Acute Pancreatitis of Rats

Objective: To investigate the effect of recombinant staphylokinase (r-Sak) and the Chinese medicine Yihuo Qingyi Decoction (益活凊胰汤 Herbal decoction for severe acute pancreatitis) in the treatment of the severe acute pancreatitis (SAP) in rats, and to observe the synergistic effect of the two. Methods: One hundred and sixty-two adult male SD rats with the body mass of 250–280 g were randomly divided into the following 5 groups: sham operation group (n=18), control group (n=36), Yihuo Qingyi Decoction treatment group (n=36), r-Sak treatment group (n=36), and Yihuo Qingyi Decoction plus r-Sak treatment group (n=36). The SAP ratmodel was prepared by retrograde injection of 5% sodium taurocholate into the cholangiopancreatic duct. Two days before modeling, Yihuo Qingyi Decoction was intragastrically administrated, and r-Sak was intraperitoneally injected. The survival rate within 18 h after modeling was determined. The pancreatic blood flow, the weight of ascites, and the serum amylase and lipase were investigated at 6 h, 12 h, and 18kh after modeling, and the pancreatic tissue was examined under light microscopy to see its pathological change. Results: The 18 h survival count of group A,B,C,D and E rats was 9,2,6,7 and 8 respectively. After r-Sak and Yihuo Qingyi Decoction intervention, the serum amylase and lipase and the weight of ascites were significantly decreased, especially in group E.18 h after modeling, the level of the serum amylase and lipase and the weight of ascites in group E was 1 100±118 U·L-1,1 000±150 U·L-1 and 13.40±1.80 g respectively, obviously lower than that of group B (P<0.05).After SAP was induced, the pancreatic blood flow showed a tendency to decrease, but the decrease extent in the treatment groups was smaller than that in the control group.18h after modeling, the pancreatic blood flow in group B and group E was 30.16±8.96 mL·100 g-1·min-1,and 129.10±42.58 mL·100 g-1·min-1 respectively, there was significant difference (P<0.05). The pathological change of the pancreatic tissue was alleviated in the treatment groups. Conclusion: Both r-Sak and Yihuo Qingyi Decoction play a beneficial role in the treatment of rat SAP and there is a synergistic effect between the two.

Study on chemical constituents and biological activities of Sanghuangporus vaninii under static magnetic field

The chemical composition and biological activity of Sanghuangporus vaninii solid culture treated by static magnetic field were studied.The compounds were isolated and purified using ultrasonic extraction and semi-preparative liquid chromatography.Their structures were identified through spectral data,and the inhibitory activities againstα-amylase,α-glucosidase and pancreatic lipase were evaluated.Three compounds,5,7-dihydroxy-3,4'-dimethoxyflavone(1),D-(+)-Trehalose(2),pinolenic acid(3),were extracted from the petroleum ether layer.Comparison of peak heights indicated a significantly higher content of compounds subjected to a 4 mT static magnetic field compared to those untreated.Activity tests revealed that compounds 1 and 2 exhibited strong anti-α-amylase and anti-α-glucosidase activities.Specifically,compound 1 had inhibition rates of(65.37±0.05)%and(73.81±0.12)%after 4 mT treatment,compared to(57.26±0.11)%and(65.33±0.14)%without.Compound 2 showed inhibition rates of(68.61±0.12)%and(65.38±0.09)%with the magnetic field,versus(60.71±0.06)%and(56.18±0.02)%without.Compound 3 displayed a notable inhibitory effect on pancreatic lipase,with rates of(60.83±0.03)%after 4 mT treatment and(53.77±0.09)%without.This study demonstrates that the static magnetic field enhances the chemical content and bioactivity of Sanghuangporus vaninii solid culture,providing a basis for its further development and application.