Usefulness of urinary trypsinogen-2 and trypsinogen activation peptide in acute pancreatitis:A multicenter study in Japan

BACKGROUND Rapid urinary trypsinogen-2 dipstick test and levels of urinary trypsinogen-2 and trypsinogen activation peptide(TAP) concentration have been reported as prognostic markers for the diagnosis of acute pancreatitis.AIM To reconfirm the validity of all these markers in the diagnosis of acute pancreatitis by undertaking a multi-center study in Japan.METHODS Patients with acute abdominal pain were recruited from 17 medical institutions in Japan from April 2009 to December 2012. Urinary and serum samples were collected twice, at enrollment and on the following day for measuring target markers. The diagnosis and severity assessment of acute pancreatitis were assessed based on prognostic factors and computed tomography(CT) Grade of the Japanese Ministry of Health, Labour, and Welfare criteria.RESULTS A total of 94 patients were enrolled during the study period. The trypsinogen-2 dipstick test was positive in 57 of 78 patients with acute pancreatitis(sensitivity,73.1%) and in 6 of 16 patients with abdominal pain but without any evidence of acute pancreatitis(specificity, 62.5%). The area under the curve(AUC) score of urinary trypsinogen-2 according to prognostic factors was 0.704, which was highest in all parameter. The AUC scores of urinary trypsinogen-2 and TAP according to CT Grade were 0.701 and 0.692, respectively, which shows higher than other pancreatic enzymes. The levels of urinary trypsinogen-2 and TAP were significantly higher in patients with extended extra-pancreatic inflammation as evaluated by CT Grade.CONCLUSION We reconfirmed urinary trypsinogen-2 dipstick test is useful as a marker for the diagnosis of acute pancreatitis. Urinary trypsinogen-2 and TAP may be considered as useful markers to determine extra-pancreatic inflammation in acute pancreatitis.

Clinical value of rapid urine trypsinogen-2 test strip, urinary trypsinogen activation peptide, and serum and urinary activation peptide of carboxypeptidase B in acute pancreatitis

AIM: To assess the usefulness of urinary trypsinogen-2 test strip, urinary trypsinogen activation peptide (TAP),and serum and urine concentrations of the activation peptide of carboxypeptidase B (CAPAP) in the diagnosisof acute pancreatitis.METHODS: Patients with acute abdominal pain and hospitalized within 24 h after the onset of symptoms were prospectively studied. Urinary trypsinogen-2 was considered positive when a clear blue line was observed (detection limit 50 μg/L). Urinary TAP was measured using a quantitative solid-phase ELISA, and serum and urinary CAPAP by a radioimmunoassay method.RESULTS: Acute abdominal pain was due to acute pancreatitis in 50 patients and turned out to be extrapancreatic in origin in 22 patients. Patients with acute pancreatitis showed significantly higher median levels of serum and urinary CAPAP levels, as well as amylase and lipase than extrapancreatic controls. Median TAP levels were similar in both groups. The urinary trypsinogen-2 test strip was positive in 68% of patients with acute pancreatitis and 13.6% in extrapancreatic controls (P<0.01). Urinary CAPAP was the most reliable test for the diagnosis of acute pancreatitis (sensitivity 66.7%, specificity 95.5%, positive and negative predictive values 96.6% and 56.7%, respectively), with a 14.6 positive likelihood ratio for a cut-off value of 2.32 nmol/L.CONCLUSION: In patients with acute abdominal pain,hospitalized within 24 h of symptom onset, CAPAP in serum and urine was a reliable diagnostic marker of acute pancreatitis. Urinary trypsinogen-2 test strip showed a clinical value similar to amylase and lipase.Urinary TAP was not a useful screening test for the diagnosis of acute pancreatitis.

Prediction of the severity of acute pancreatitis on admission by urinary trypsinogen activation peptide: A meta-analysis

AIM: To undertake a meta-analysis on the value of urinary trypsinogen activation peptide (uTAP) in predicting severity of acute pancreatitis on admission.METHODS: Major databases including Medline, Embase, Science Citation Index Expanded and the Cochrane Central Register of Controlled Trials in the Cochrane Library were searched to identify all relevant studies from January 1990 to January 2013. Pooled sensitivity, specificity and the diagnostic odds ratios (DORs) with 95%CI were calculated for each study and were compared to other systems/biomarkers if mentioned within the same study. Summary receiver-operating curves were conducted and the area under the curve (AUC) was evaluated.RESULTS: In total, six studies of uTAP with a cut-off value of 35 nmol/L were included in this meta-analysis. Overall, the pooled sensitivity and specificity of uTAP for predicting severity of acute pancreatitis, at time of admission, was 71% and 75%, respectively (AUC = 0.83, DOR = 8.67, 95%CI: 3.70-20.33). When uTAP was compared with plasma C-reactive protein, the pooled sensitivity, specificity, AUC and DOR were 0.64 vs 0.67, 0.77 vs 0.75, 0.82 vs 0.79 and 6.27 vs 6.32, respectively. Similarly, the pooled sensitivity, specificity, AUC and DOR of uTAP vs Acute Physiology and Chronic Health Evaluation II within the first 48 h of admission were found to be 0.64 vs 0.69, 0.77 vs 0.61, 0.82 vs 0.73 and 6.27 vs 4.61, respectively.CONCLUSION: uTAP has the potential to act as a stratification marker on admission for differentiating disease severity of acute pancreatitis.

Urinary trypsinogen-2 for diagnosing acute pancreatitis:a meta-analysis

BACKGROUND: Currently, serum amylase and lipase are the most popular laboratory markers for early diagnosis of acute pancreatitis with reasonable sensitivity and specificity. Urinary trypsinogen-2 (UT-2) has been increasingly used but its clinical value for the diagnosis of acute pancreatitis and post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis has not yet been systematically assessed. DATA SOURCES: A comprehensive search was carried out using PubMed (MEDLINE), Embase, and Web of Science for clinical trials, which studied the usefulness of UT-2 as a diagnostic marker for acute pancreatitis. Sensitivity, specificity and the diagnostic odds ratios (DORs) with 95% confidence interval (CI) were calculated for each study and were compared with serum amylase and lipase. Summary receiver-operating curves were conducted and the area under the curve (AUC) was evaluated. RESULTS: A total of 18 studies were included. The pooled sensitivity and specificity of UT-2 for the diagnosis of acute pancreatitis were 80% and 92%, respectively (AUC=0.96, DOR=65.63, 95% CI: 31.65-139.09). The diagnostic value of UT-2 was comparable to serum amylase but was weaker than serum lipase. The pooled sensitivity and specificity for the diagnosis of post-ERCP pancreatitis were 86% and 94%, respectively (AUC=0.92, DOR=77.68, 95% CI: 24.99-241.48).CONCLUSIONS: UT-2 as a rapid test could be potentially used for the diagnosis of post-ERCP pancreatitis and to an extent, acute pancreatitis. Further studies are warranted to confirm these results.

Early prediction of severe acute pancreatitis by urinary trypsinogen activation peptide

Objective: To investigate the value of urinary trypsi-nogen activation peptide (TAP) in the early predic-tion of severe acute pancreatitis and to compare itwith acute physiology and chronic health evaluationⅡ (APACHE Ⅱ).Methods: We assessed the predictive value of urinaryTAP concentrations measured by a competitive en-zyme-linked immunosorbent assay. Urine sampleswere collected for detecting TAP concentrations atadmission, and 24, 48, and 72 h from 41 patientswith acute pancreatitis (12 with severe disease, 29with mild disease) who presented within 48 h the on-set of symptoms and from 11 control patients, whileAPACHE Ⅱ scores were recorded at 48 h after ad-mission.Results: The peak median urinary TAP concentrationwas seen at admission. The median urinary TAPconcentration at admission for severe pancreatitis (95nmol/L) was significantly higher than the medianfor patients with mild pancreatitis (20 nmol/L, P【0. 005) and controls (15 nmol/L, P【0. 005). TAPconcentrations were significantly higher in patientswith severe acute pancreatitis than the median in pa-tients with mild pancreatitis (P【0. 05) and controls(P【0. 05) on days 2 to 3. The median APACHE Ⅱscores of severe patients were significantly differentfrom those of mild patients (10.5 vs 6.0, P【0.01).The sensitivity, specificity, positive predictive, andnegative predictive values of an admission urinaryTAP≥35 nmol/L for severe pancreatitis were91.7%, 89.7%, 78.6% and 96.3%, whereas 48 hafter admission the values for APACHE Ⅱ scores(≥9) were 75.0%, 72.7%, 52.9% and 87.5%. Inprediction of disease severity, the urine TAP concen-tration was much better than APACHE Ⅱ at 48 h.Conclusions: Urinary TAP obtained at the first 48 hof the onset of symptoms can predict severe acutepancreatitis. In prediction of disease severity, theurinary TAP is much better than APACHE Ⅱ score.

Relationship between Carbachol Hyperstimulation-Induced Pancreatic Acinar Cellular Injury and Trypsinogen or NF-κB Activation in Rats in vitro

The relationship between M3 cholinergic receptor agonist （carbachol） hyperstimulationinduced pancreatic acinar cellular injury and trypsinogen activation or NF-κB activation in rats was studied in vitro. Rat pancreatic acinar ceils were isolated, cultured and treated with carbachol, the active protease inhibitor （pefabloc）, and NF-κB inhibitor （PDTC） in vitro. Intracellular trypsin activity was measured by using a fluorogenic substrate. The cellular injury was evaluated by measuring the leakage of LDH from pancreatic acinar ceils. The results showed that as compared with control group, 10-3 mol/L carbachol induced a significant increase of the intracellular trypsin activity and the leakage of LDH from pancreatic acinar cells. Pretreatment with 2 mmol/L pefabloc could significantly decrease the activity of trypsin and the leakage of LDH from pancreatic acinar cells （P〈0. 01） following the treatment with a high concentration of carbachol （10^-3 mol/L） in vitro. The addition of 10^-2mol/L PDTC didn＇t result in a significant decrease in the activity of trypsin and the leakage of LDH from pancreatic acinar cells treated with a high concentration of carbachol （10^-3 mol/L） in vitro （P〉0. 05）. It was concluded that intracellular trypsinogen activation is likely involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro. NF-κB activation may not be involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro.

Relationship between Carbachol Hyperstimulation-induced Pancre-atic Intracelluar Trypsinogen and NF-κB Activation in Rats in vitro

The relationship between intracelluar trypsinogen activation and NF-κB activation in rat pancreatic acinar cells induced by M3 cholinergic receptor agonist （carbachol） hyperstimulation was studied. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, the active protease inhibitor （pefabloc） and NF-κB inhibitor （PDTC） in vitro. Intracelluar trypsin activity was measured by using a fluorogenic substrate. The activity of NF-κB was monitored by using electrophoretic mobility shift assay. The results showed that after pretreatment with 2 mmol/L pefabloc, the activities of trypsin and NF-κB in pancreatic acinar cells treated with high concertrations of carbachol （10^-3 mol/L） in vitro was significantly decreased as compared with control group （P〈0.01 ）. The addition of 10^-2 mol/L PDTC resulted in a significant decrease of NF-κB activities in pancreatic acinar cells after treated with high concertrations of carbachol （10^-3 mol/L） in vitro, but the intracelluar trypsinogen activity was not obviously inhibited （P〉0.05）. It was concluded that intracelluar trypsinogen activation is likely involved in the regulation of high concertrations of carbachol-induced NF-κB activation in pancreatic acinar cells in vitro. NF-κB activation is likely not necessary for high concertrations of carbachol-induced trypsinogen activation in pancreatic acinar cells in vitro.

Use of the urinary trypsinogen-2 dipstick test in early diagnosis of pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP)

Background: Acute pancreatitis is one of the most serious complications of ERCP. Early diagnosis of post ERCP pancreatitis helps physicians to provide intensive care and possible medical treatment as early as possible. Trypsinogen-2 in urine is a good diagnostic and prognostic marker of acute pancreatitis. Objectives: To evaluate the diagnostic value of urinary trypsinogen-2 dipstick test for early diagnosis of post ERCP pancreatitis. Methods: A total of 37 patients with obstructive jaundice were tested with the urinary trypsinogen-2 dipstick test and serum levels of amylase and lipase before ERCP and 6 hours after ERCP. Results: Post ERCP pancreatitis was diagnosed in 6 (16%) of 37 patients. The sensitivity, specificity, positive predictive value and negative predictive value of urinary trypsinogen-2 dipstick test at 6 hours after ERCP were 100%, 97%, 86%, 100% respectively. At the cutoff level (130 U/L) for lipase, the positive predictive value and negative predictive value all were (100%), however, the positive predictive value and negative predictive value for amylase levels at cutoff (122 U/L) were 60%, 100% respectively. Serum lipase level was the best test for diagnosing post ERCP pancreatitis followed by the urinary trypsinogen-2 dipstick test. Conclusions: The urinary trypsinogen-2 dipstick test can be used as a rapid and easy test for early diagnosis of post ERCP pancreatitis with high sensitivity and specificity.

PRSS1 c.455-33C>T突变在遗传性胰腺炎中的致病性分析

目的探讨人阳离子胰蛋白酶原(PRSS1)c.455-33C>T内含子杂合突变在遗传性胰腺炎(HP)中的致病性。方法报告1例反复发作的急性胰腺炎病人,病人携带PRSS1 c.455-33C>T杂合突变,检索文献及网站初步分析其突变。提取健康对照者外周血基因组DNA,通过PCR扩增将PRSS1基因3号内含子序列、4号外显子序列和4号内含子序列与pSPL3载体连接构建质粒,命名为SWT;在SWT质粒的基础上通过定点突变的方法构建PRSS1 c.455-33C>T突变质粒,命名为S33;转染人胚肾细胞(Hek293T)后提取总RNA并进行RT-PCR及凝胶电泳,分析PRSS1 c.455-33C>T内含子突变是否引起PRSS1 mRNA剪切的改变进而引起HP。PCR扩增健康对照者外周血基因组DNA,将PRSS1基因3号内含子序列与增强子分析质粒pGL4.23连接,命名为EWT质粒;并在EWT质粒的基础上构建PRSS1 c.455-33C>T定点突变的质粒,命名为E33;将EWT、E33质粒转染Hek293T后利用双荧光素酶报告基因分析系统进行荧光素酶活性测定,分析PRSS1 c.455-33C>T内含子突变是否通过具有增强子功能引起PRSS1基因功能发生改变进而导致HP。结果检索文献及网站确定PRSS1 c.455-33C>T突变为未报道的功能不明新突变。剪切分析实验显示,SWT和S33两组RT-PCR产物相同;双荧光素酶实验结果显示,E33组的荧光值低于EWT组,差异有统计学意义(t=12.23,P<0.001),E33组与EWT组相比没有增强子功能。结论PRSS1 c.455-33C>T内含子突变不引起PRSS1 mRNA剪切的改变,也不具有PRSS1基因增强子功能,与PRSS1基因直接导致的HP无关。

Uamy/Ucr比值及尿胰蛋白酶原-2水平评估急性胰腺炎患者预后的临床价值

目的探讨尿淀粉酶/尿肌酐(Uamy/Ucr)比值及尿胰蛋白酶原-2(TPS-2)水平对急性胰腺炎(AP)患者预后的评估价值。方法选取2019年3月—2021年5月在成都市第三人民医院收治的104例AP患者,根据其预后情况分为存活组(n=71)和死亡组(n=33)。比较两组患者的临床资料,采用化学比色法检测血清钙(Ca^(2+))水平,采用动脉血气分析仪检测动脉血氧分压(PaO_(2))水平,采用酶联免疫法检测Uamy和Ucr水平,采用免疫层析法检测尿TPS-2水平,应用受试者工作特征(ROC)曲线分析Uamy/Ucr比值和尿TPS-2对AP患者预后的预测价值。结果死亡组患者年龄、体质指数(BMI)、合并高血脂、Ca^(2+)和PaO_(2)水平与存活组比较差异有统计学意义(P<0.05);死亡组患者Uamy和Ucr均高于存活组(P<0.05);死亡组患者Uamy/Ucr比值和尿TPS-2阳性率分别为90.91%、93.94%,高于存活组的56.92%、53.52%(P<0.05)。ROC曲线分析显示,Uamy/Ucr比值、尿TPS-2及联合检测预测AP患者死亡的曲线下面积(AUC)分别为0.694、0.702、0.858(P<0.05)。结论AP患者预后与其年龄、血清Ca^(2+)和PaO_(2)水平有关。Uamy/Ucr比值及TPS-2水平对AP患者预后有较高的预测价值。

DKK-1、TAT-2联合血清胃蛋白酶原、胃泌素-17在胃癌早期诊断中的临床价值分析

目的 探讨血清Dickkopf相关蛋白1(DKK-1)、肿瘤相关胰蛋白酶原-2(TAT-2)联合胃蛋白酶原(PG)、胃泌素-17(G-17)在胃癌早期诊断中的临床价值。方法 选取2021年10月至2022年10月在该院行胃镜检查发现胃黏膜病变的患者125例作为研究对象,其中癌前病变31例(癌前病变组)、早期胃癌32例(早期胃癌组)、进展期胃癌32例(进展期胃癌组)、慢性非萎缩性胃炎30例(对照组)。检测所有患者血清中DKK-1、TAT-2、PG及G-17水平,采用受试者工作特征(ROC)曲线探讨其在胃癌早期诊断中的临床价值。结果 与对照组比较,癌前病变组、早期胃癌组、进展期胃癌组PG-Ⅰ水平逐渐降低,PG-Ⅱ、G-17、DKK-1、TAT-2水平逐渐升高,差异均有统计学意义(P<0.05),但早期胃癌组与进展期胃癌组TAT-2水平比较,差异无统计学意义(P>0.05)。血清DKK-1水平与胃癌患者肿瘤最大径、淋巴结转移、分化程度及临床分期有关(P<0.05)。ROC曲线分析结果显示,PG-Ⅰ、PG-Ⅱ、G-17、DKK-1、TAT-2诊断胃癌的曲线下面积(AUC)分别为0.726、0.862、0.789、0.870、0.914(P<0.05),新ABC法+DKK-1、新ABC法+TAT-2诊断胃癌的AUC分别为0.979、0.980(P<0.05),所有指标联合诊断胃癌的AUC为0.993。结论 PG、G-17作为一种传统的血清学标志物,诊断价值确切,DKK-1、TAT-2水平随胃黏膜病变的不同表现差异明显,上述所有指标联合诊断对胃癌有较高的价值。

一个遗传性胰腺炎家系中新发现的胰蛋白酶原基因突变

对1个遗传性胰腺炎(hereditary pancreatitis,HP)家系中6例成员和120例无亲缘关系健康人的胰蛋白酶原基因(protease serine 1,PRSS1)进行PCR扩增,产物纯化后测序,结合受检者的血清肿瘤标志物、糖尿病相关生化指标以及近亲属的一般临床资料进行分析。结果发现4例家系成员PRSS1基因3号外显子区136位碱基存在C→T杂合性突变,他们的基因型表现为野生型与突变型杂合现象,另外在先证者PRSS1基因的3号外显子区171位碱基还存在着一个同义突变点(C→T),而对照组和家系其他成员中未发现此两种突变,突变阳性患者表现为乳酸、糖基化血红蛋白和糖类肿瘤标志物(CA19-9、CA125)增高。因此,PRSS1基因3号外显子区136位碱基C→T杂合性突变与该家系遗传性胰腺炎有关,是该家系中遗传性胰腺炎的遗传易感因素。

3种益生素配伍对异育银鲫(Carassius auratus gibelio)生长、消化及肠道菌群组成的影响

选用196尾1龄异育银鲫[体重为(30±2)g],随机分成4组,其中,1组为对照组,2、3、4为试验组,1组投喂的为基础饲料,2、3、4组投喂的饲料是在基础饲料中分别添加100μg/g地衣芽孢杆菌、100μg/g地衣芽孢杆菌与100μg/g低聚木糖、100μg/g地衣芽孢杆菌与300μg/g复合酶制剂,在室内流水养殖系统中饲养112天。研究3种益生素及其配伍对异育银鲫生长、消化及肠道菌群组成等的影响。结果表明,2、3、4组异育银鲫的增重率比1组(对照)均有提高,并分别比对照组提高了23.97%、43.78%和18.36%(P<0.05)。试验组的营养物质表观消化率、肠道酶活性及肝胰脏蛋白酶mRNA表达量与对照组相比均有提高且差异显著(P<0.05)。在肠道菌群组成上,各试验组大肠杆菌数和乳酸杆菌均比对照组少且差异显著(P<0.05);而在肠道芽孢杆菌方面,除芽孢杆菌与酶制剂配伍组外,其余两试验组对芽孢杆菌均有促生长作用,芽孢杆菌与寡糖配伍组对芽孢杆菌的促生长作用明显(P<0.05)。由此可见,芽孢杆菌、低聚木糖、复合酶制剂及它们的配伍物可以促进异育银鲫的生长,提高饲料利用率,促进肠道有益微生物的生长和抑制有害微生物,并且能提高肝胰脏蛋白酶mRNA表达量和肠道酶活性。

斑马鱼3种胰蛋白酶原基因的克隆、序列分析及组织表达

胰蛋白酶是丝氨酸蛋白酶类超家族成员之一,在动物蛋白消化中起着重要作用。为深入研究胰蛋白酶在鱼类中的蛋白结构和生理功能,利用RT-PCR和RACE方法,成功获得了斑马鱼3种胰蛋白酶原cDNA序列(zftry1a、zftry1b和zftry2)。结果表明,zftry1a和zftry1b均有242个氨基酸残基组成,其中包括15个氨基酸的信号肽和5个氨基酸(LDDDK)的激活肽。zftry2由247个氨基酸残基组成,其中包括15个氨基酸的信号肽和9个氨基酸(APLGDDDDK)的激活肽。氨基酸序列比对结果显示,三者具备胰蛋白酶原的保守结构特征,如含有催化三联体氨基酸(His-57、Asp-102和Ser-195),12个半胱氨酸,位于底物结合口袋底部Asp-189和口袋开口处的Gly-216、Gly-226等。进化树结果显示,斑马鱼zftry1a和zftry1b属于group I,为阴离子胰蛋白酶原;斑马鱼zftry2属于group Ⅱ,为阳离子型胰蛋白酶原。RT-PCR结果显示,三者组织分布模式类似,且在肠中有最高表达量。这些结果为研究鱼类胰蛋白酶原的基因进化和功能以及进一步探讨鱼类消化生理的分子机制奠定了基础。

斜带石斑鱼胰蛋白酶原和淀粉酶全长cDNA的克隆与序列分析

采用RT-PCR及RACE法从斜带石斑鱼Epinephelus coioides肝胰脏克隆得到胰蛋白酶原(trypsinogen,TRY)与淀粉酶(amylase,AMY)基因cDNA全序列。斜带石斑鱼肝胰脏TRY基因cDNA全长911bp,其中5'非翻译区(5'-UTR)为55bp,3'-UTR为127bp,开放阅读框(ORF)为729bp,编码242个氨基酸,包含所有丝氨酸蛋白酶中共有的高度保守的催化活性中心。序列一致性分析发现,斜带石斑鱼与牙鲆Paralichthys olivaceus、金头鲷Sparus aurata、鳎Solea senegalensis、石鲽Pleuronectes bicoloratus的TRY序列相似性高达86.8%-89.7%,与人Homo sapiens、小鼠Mus musculus、斑马鱼Danio rerio的TRY相似性较低为59.9%-64.5%。斜带石斑鱼AMY基因cDNA全长1657bp,其中5'-UTR为41bp,3'-UTR为77bp,ORF为1539bp,编码512个氨基酸,包含与哺乳动物α-AMY二级结构相似的8个α螺旋和8个β折叠。序列一致性分析发现,斜带石斑鱼与澳洲肺鱼Neoceratodus forsteri、美洲拟鲽Limanda americanus、大西洋鲑Salmo salar、斑马鱼AMY基因序列相似性高达82.4%-91.8%,与人、小鼠、鸡G.Gallus的AMY基因相似性较低为70.1%-72.3%。斜带石斑鱼TRY和AMY基因cDNA全序列的成功克隆为进一步研究其表达调控机理及研发有效提高其表达水平的饲料添加剂奠定基础。

生化检测指标在急性胰腺炎诊断中的临床意义

目的探讨血淀粉酶(S-Amy)、尿淀粉酶(U-Amy)、胰脂肪酶(LPS)、胰蛋白酶原激活肽(TAP)和C反应蛋白(CRP)等5项生化指标在急性胰腺炎(AP)早期诊断和病情评估中的应用价值。方法将265例急腹症患者分为AP组和非AP组,其中AP组根据病情严重程度分为轻症AP(MAP)和重症AP(SAP),同时选取35例健康体检者作为正常对照组。检测各组5项指标。S-Amy、U-Amy和LPS采用速率法检测,TAP采用双抗体夹心酶联免疫吸附试验(ELISA)检测,CRP采用乳胶增强速率散射比浊法检测。结果 S-Amy、U-Amy和TAP水平由高到低依次为AP组>非AP组>正常对照组,各组间差异均有统计学意义(P<0.01)。AP组LPS水平明显高于非AP组和正常对照组(P<0.01),AP组和非AP组CRP水平均高于正常对照组(P<0.01)。SAP组U-Amy、TAP及CRP水平明显高于MAP组(P<0.01),而S-Amy、LPS水平2组间无差异(P>0.05)。在早期AP的诊断中,血清LPS敏感性为92.1%、特异性为91.6%,明显高于S-Amy、U-Amy、TAP和CRP。结论 LPS的检测有助于AP的早期诊断,而TAP和CRP则有助于病变严重程度的判断以及治疗效果的观察与预后评估。

胰蛋白酶原激活肽和白细胞介素-6在实验性急性胰腺炎中的意义

目的探讨血浆胰蛋白酶原激活肽(TAP)和血清白细胞介素-6(IL-6)作为早期预测实验性急性胰腺炎病情严重程度指标的可行性。方法雄性SD大鼠90只,随机分成5组。分别是3%牛磺胆酸钠逆行胆胰管注射组、5%牛磺胆酸钠逆行胆胰管注射组、3%牛磺胆酸钠逆行胆胰管注射后0.5h经股静脉注入乌司他丁组、0.9%生理盐水逆行胆胰管注射组和假手术组(只行开、关腹术)。每组18只大鼠,每组再随机分成3组,分别于制模后3、6、24 h后取血处死,留取标本。观察血清淀粉酶、血浆TAP水平、血清IL-6水平变化,并在光镜下对胰腺病变进行双盲组织病理学评分。结果制模后3和6 h血浆TAP水平5%牛磺胆酸钠逆行胆胰管注射组[(4.798±0.169)和(3.999±0.299)nmol/L]比3%牛磺胆酸钠逆行胆胰管注射组[(2.416±0.148)和(3.356±0.211)nmol/L]明显增高(P<0.01);制模后6 h,血清IL-6水平5%牛磺胆酸钠逆行胆胰管注射组(1339.51±56.43)pg/ml明显高于3%牛磺胆酸钠逆行胆胰管注射组(619.07±42.25)pg/ml(P<0.01)。结论在轻重不同的急性胰腺炎模型中,胰腺病理改变越严重,血浆TAP和血清IL-6峰值出现越早。血浆TAP水平可以作为早期精确预测急性胰腺炎模型胰腺病变严重程度的指标。

蛋白质分子量测定过程中的酸效应

在基质辅助激光解吸电离飞行时间质谱 (MALDI TOF MS)和电喷雾质谱 (ESI MS)测定蛋白质分子分子量的过程中 ,适当提高样品的酸度 ,可提高分析测试的灵敏度。在选定最佳样品分子浓度的基础上 ,通过适当加入三氟乙酸 (TFA)来调整测试样品的酸度 ,准确测定了标准蛋白质 溶菌酶 (lysozyme)的分子量 ,并且对蛋白质分子在“软电离”质谱中 。

诊断急性胰腺炎的生化检测指标评析

目的通过检测血淀粉酶(S-Amy)、胰脂肪酶(LPS)、C反应蛋白(CRP)及尿淀粉酶(U-Amy)、尿胰蛋白酶原激活肽(TAP)5项生化指标,探讨急性胰腺炎(AP)早期诊断及病情评估的有效指标。方法将541例急腹症患者分为AP组和非AP组,其中AP组分为轻症AP(MAP)和重症AP(SAP),同时选取80例健康体检者作为正常对照组,于患者入院后6h内采集静脉血并留取新鲜尿液,对各组5项生化指标进行检测和分析。S-Amy、U-Amy、LPS采用速率法,TAP、CRP采用ELISA法。结果 S-Amy、U-Amy和TAP水平在AP组最高,非AP组次之,正常对照组最低,各组之间差异均具有统计学意义(P<0.01)。AP组LPS水平较非AP组及正常对照组显著增高(P<0.01)。AP组与非AP组CRP水平均较正常对照组增高(P<0.01),但AP组CRP水平与非AP组比较无显著差异。SAP组U-Amy、TAP及CRP水平显著高于MAP组(P<0.01),而S-Amy、LPS水平在这两组间无显著差异(P>0.05)。LPS用于AP早期诊断的敏感度为92.1%、特异度为91.6%,显著高于S-Amy、U-Amy、TAP及CRP。结论血、尿淀粉酶、LPS指标对于AP的早期诊断具有重要意义,TAP和CRP有助于评估病变的严重程度、观察疗效及评估预后。

血清淀粉酶、血清胰蛋白酶原-2、血小板活化因子及IL-10在ERCP术后胰腺炎患者中的临床价值研究

目的：探讨血清淀粉酶、血清胰蛋白酶原-2、血小板活化因子、IL-10在ERCP术后胰腺炎患者中的临床价值。方法：选取本院消化内科住院的ERCP患者256例作为研究对象,按照ERCP术后不同的症状、体征、血清淀粉酶和胰腺CT检查结果,将研究对象分为对照组、高淀粉酶血症组及PEP组3组,检测其血清淀粉酶、血清胰蛋白酶原-2、血小板活化因子、IL-10水平并进行比较。结果 ：3组血清淀粉酶、血清胰蛋白酶原-2、血小板活化因子、IL-10比较,差异均有统计学意义（F分组=269.578、139.492、145.637、126.891,均P=0.000）,PEP组均为最高值。各个时间点的血清淀粉酶、血清胰蛋白酶原-2、血小板活化因子、IL-10比较,差异也有统计学意义（F时间=602.506、494.375、462.512、239.564,均P=0.000）,高淀粉酶血症组的血清淀粉酶术后4h达到最高值,术后24h已经降低但仍高于正常水平,PEP组的血清淀粉酶术后4h急剧升高,到术后24h仍在升高;高淀粉酶血症组及PEP组血清胰蛋白酶原-2、血小板活化因子、IL-10均在术后4h达到最高值且术后24h均降低。PEP组血清淀粉酶、血清胰蛋白酶原-2、血小板活化因子、IL-10术后4h、术后24h较术前的增高幅度均高于高淀粉酶血症组及对照组（F交互=28.492、22.614、16.573、7.819,均P=0.000）。PEP组血清淀粉酶与血清胰蛋白酶原-2、血小板活化因子、IL-10均有正相关关系（r=0.591、0.662、0.681,均P〈0.05）。结论 ：血清淀粉酶、血清胰蛋白酶原-2、血小板活化因子、IL-10在ERCP术后胰腺炎患者中具有重要的临床诊断价值。