Differences in Newborn Screening Results Among Women with Gestational Diabetes Mellitus

Abstract Multiple studies undertaken on cord blood demonstrate analyte perturbations in infants exposed to gestational diabetes mellitus (GDM). Cord blood as a sample is influenced by maternal and placental metabolism. Newborn screening (NBS), performed after the first 24 hours of life reflects early neonatal metabolism. We compared NBS analytes between women with and without GDM with different management approaches in the Treatment of Booking of Gestational Diabetes (TOBOGM) pilot randomised controlled trial. Pregnant women with GDM risk factors were randomised to early or deferred GDM treatment following an oral glucose tolerance test (<20 weeks gestation). Women without GDM served as "decoys". From the decoy group 11 developed GDM (screened at 26-28 weeks), were analysed separately; their results were compared with the other groups. De-identified controls were chosen from NBS results from the same analytic run matched for sex, birthweight and gestational age. Results were available for 73/78 women participating in the pilot and 358 de-identified controls. Tyrosine levels (μmol/l; whole blood)were higher in the late GDM group vs early, deferred treatment, and decoy groups (medians:106.28; IQR: 96.73-151.11) (76.33; 64.64-97.90) (75.68; 66.59-110.88)(73.74; 58.32-90.36) (p=0.009) and remained elevated when compared to normal, age-matched controls (106.28; 96.73-151.11) (87.26; 68.55-111.26) (p value=0.01) Immunoreactive trypsinogen (μgm/l; whole blood)was highest in the early treatment group when compared with group-specific controls (22.30; 13.90-29.90 vs 14.00, 10.60-21.10) (p=0.02). These results provide evidence of biochemical perturbations detectable on NBS of in-utero exposure to hyperglycemia and treatment and provide data for hypothesis building.

The first five-year evaluation of cystic fibrosis neonatal screening program in São Paulo State, Brazil / Uma avaliação dos primeiros cinco anos do programa de triagem neonatal da fibrose cística no Estado de São Paulo, Brasil / Una evaluación de los primeros cinco años del programa de cribado neonatal de fibrosis cística en el Estado de São Paulo, Brasil

The Hospital of the Ribeirão Preto Medical School, University of São Paulo is one of the three screening centers in São Paulo State, Brazil, and has included a test for cystic fibrosis (CF) since February 6, 2010, by a court order. We evaluated the first five years of this CF-newborn screening program. The original immunoreactive trypsinogen (IRT)/IRT screening protocol was adopted in Brazil. A total of 173,571 newborns were screened, 1,922 (1.1%) of whom showed IRT1 ≥ 70ng/mL. Of these, 1,795 (93.4%) collected IRT2, with elevated results (IRT2 ≥ 70ng/mL) in 102 of them (5.2%). We identified a total of 26 CF cases during this period, including three CF cases that were not detected by the CF-newborn screening. The incidence of the disease among the screened babies was 1:6,675 newborns screened. Median age at the initial evaluation was 42 days, comparable to that of neonates screened with the IRT/DNA protocol. Almost all infants with CF already exhibited some manifestations of the disease during the neonatal period. The mutation most frequently detected in the CF cases was F508del. These findings suggest the early age at the beginning of treatment at our center was due to the effort of the persons involved in the program regarding an effective active search. Considering the false negative results of CF-newborn screening and the early onset of clinical manifestations of the disease in this study, pediatricians should be aware of the diagnosis of CF even in children with negative test.

O Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo é um dos três centros de triagem da fibrose cística (FC) no estado de São Paulo, tendo incluído esse teste desde 6 de fevereiro de 2010, a partir de uma liminar judicial. O estudo avalia os primeiros cinco anos desse programa de triagem neonatal da FC. O Brasil adota o protocolo de triagem original, com o tripsinogênio imunorreativo (IRT)/IRT. Foram triados um total de 173.571 recém-nascidos, dos quais 1.922 (1,1%) mostraram IRT ≥ 70ng/mL. Destes, 1.795 (93,4%) tiveram amostras coletadas para IRT2, com resultados elevados (IRT2 ≥ 70ng/mL) em 102 deles (5,2%). Identificamos um total de 26 casos de FC durante esse período, inclusive 3 casos de FC que não foram detectados com a triagem neonatal. A incidência da FC foi de 1 caso em cada 6.675 recém-nascidos triados. A idade mediana na avaliação inicial foi 42 dias, comparável à idade de recém-nascidos triados com o protocolo IRT/DNA. Quase todos os lactentes com FC já exibiam algumas manifestações da doença durante o período neonatal. A mutação mais comum nos casos de FC foi a F508del. Os resultados em nosso centro indicam que a idade precoce no início do tratamento foi devido aos esforços do programa na implementação de uma busca ativa eficaz. Considerando os resultados falsos-negativos no programa de triagem neonatal para FC e o início precoce das manifestações clínicas da doença neste estudo, os pediatras devem estar cientes da possibilidade de diagnóstico de FC, mesmo em crianças com teste negativo.

El Hospital das Clínicas de la Facultad de Medicina de Ribeirão Preto, São Paulo Universidad es uno de los tres centros de cribado de fibrosis cística (FC) en el estado de São Paulo, incluyendo este test desde el 6 de febrero de 2010, debido a una medida cautelar judicial. El estudio evalúa los primeros cinco años de este programa de cribado neonatal de FC. Brasil adopta el protocolo de cribado original, con el tripsinógeno inmunorreactivo (TIR)/IRT. Se cribaron un total de 173.571 recién nacidos, de los cuales 1.922 (1,1%) mostraron IRT ≥ 70ng/mL. De estos, se obtuvieron 1.795 (93,4%) muestras recogidas para IRT2, con resultados elevados (IRT2 ≥ 70ng/mL) en 102 de ellos (5,2%). Identificamos un total de 26 casos de FC durante ese período, inclusive 3 casos de FC que no fueron detectados con el cribado neonatal. La incidencia de la FC fue de 1 caso por cada 6.675 recién-nacidos cribados. La edad media en la evaluación inicial fue 42 días, comparable a la edad de recién nacidos cribados con el protocolo IRT/DNA. Casi todos los lactantes con FC ya manifestaban algunos síntomas de la enfermedad durante el período neonatal. La mutación más común en los casos de FC era el F508del. Los resultados en nuestro centro indican que la edad precoz en el inicio del tratamiento se debía a los esfuerzos del programa en la implementación de una búsqueda activa eficaz. Considerando los resultados falsos-negativos en el programa de cribado neonatal para FC, y el inicio precoz de las manifestaciones clínicas de la enfermedad en este estudio, los pediatras deben ser conscientes de la posibilidad de diagnóstico de FC, incluso en niños con test negativo.

Urinary trypsinogen-2 dipstick test for point-of-care screening of acute pancreatitis

Background: Acute pancreatitis (AP) is associated with high mortality in its severe form. Conventional laboratory tests used in its diagnosis are fraught with multiple shortcomings. Early institution of intravenous fluid resuscitation can reduce morbidity and mortality. Measurement of urinary trypsinogen-2 using a bedside urine dipstick test may prove useful in early identification of AP.Methods: Patients with symptoms consistent with AP, attending the emergency department, at a tertiary care hospital in southern India, between November 2014 and November 2016, were included in a prospective observational study. The patients underwent routine investigations and additionally were tested with a urinary trypsinogen-2 dipstick test (UTT). The diagnostic performance and the time to reporting of the different investigations were compared with those of UTT. Final diagnosis of AP, made by clinicians, served as the standard.Results: The sensitivities of serum amylase, serum lipase, UTT, ultrasonography (USG) and contrast-enhanced computed tomography (CECT) were 97.1%, 94.1%, 92.7%, 98.3% and 100%, respectively. The respective specificities were 92.4%, 98.5%, 98.5%, 100% and 100%. The average time required to obtain the test report was about half hour from admission in case of UTT, compared to about 3 hours for serum amylase/lipase, 4 hours for USG and 6 hours for CECT.Conclusions: The results indicate that UTT test, due to its high performance indices, simplicity and faster availability of reports, can serve as an ideal screening test for AP and help in early institution of treatment.

Roles of Serum Trypsinogen-2 and Hematocrit in Evaluating Severity of Acute Pancreatitis PAN / 胃肠病学

The incidence of acute pancreatitis (AP) in China is increasing year by year, and the mortality remains high. It is of great significance to explore the blood biochemical parameters which can assess the severity of AP for the diagnosis and treatment of AP. Aims: To explore the value of serum trypsinogen-2 (TAT-2) and hematocrit (HCT) in assessing the severity of AP. Methods: A total of 175 cases of AP from September 2016 to June 2018 at Jiangyin People's Hospital were enrolled. Patients were divided into mild acute pancreatitis (MAP) group, moderate severe acute pancreatitis (MSAP) group, severe acute pancreatitis (SAP) group according to the severity of AP. And 76 healthy individuals undergoing physical examination were served as the control group. The levels of serum TAT-2, HCT were detected. BISAP score, APACHEⅡ score, Balthazar CT score were evaluated. Values of different parameters for diagnosing SAP were analyzed by ROC curve. Results: Differences in TAT-2, HCT, BISAP score, APACHEⅡ score, Balthazar CT score were statistically significant among the four groups (P<0.05). Serum TAT-2 and HCT in SAP group were significantly higher than those in MAP group, MSAP group and control group (P<0.05), serum TAT-2 and HCT in MSAP group were significantly higher than those in MAP group and control group (P<0.05), and serum TAT-2 and HCT in MAP group were significantly higher than those in control group (P<0.05). ROC curve analysis showed that the sensitivity of TAT-2, HCT and the combination of these two indicators for predicting SAP were 69.7%, 84.7%, 88.5%, respectively; specificity were 95.3%, 67.8%, 65.1%, respectively; AUC were 0.858, 0.783, 0.875, respectively. Conclusions: TAT-2 and HCT can effectively evaluate the severity of AP patients, the combined detection of the two indicators can predict the occurrence of SAP further and play a vital role in treatment of AP.

Fibrosis quística y tamizaje neonatal / Newborn screening and cystic fibrosis

Newborn screening for cystic fibrosis is a public health strategy that as been associated with early improved nutrition and survival, and the potential for preventing severe health problems. Various protocols have been employed, being the measurement of immunoreactive trypsynogen (IRT) in the first few days of life the first step in all of them. The second tier can include IRT/IRT, IRT/DNA, IRT/PAP (pancreatitis associated protein). Protocol selection depends on the priorities for each country but at present there is no optimal strategy.

El tamizaje neonatal para fibrosis quística es una estrategia de salud pública que ha demostrado beneficios nutricionales, aumento de la sobrevida y potencialmente prevención de problemas severos de salud. En el mundo se usan variados protocolos, sin embargo en todos el primer paso es la determinación de tripsinógeno inmunorreactivo (IRT) en sangre, tomado del talón del recién nacido. El segundo paso incluye la determinación de un segundo IRT o determinación de DNA o PAP (Proteína asociada a pancreatitis). La selección del protocolo a seguir depende de cada país pero hasta ahora no hay una estrategia óptima.

Pesquisa neonatal en fibrosis quística: un modelo de coordinación multidisciplinaria. ha llegado la hora de los centros de fibrosis quística / Cystic fibrosis screening: amultidisciplinary coordinationapproach. the time for cysticfibrosis centers has arrived

Introducción. La pesquisa neonatal para fibrosis quística ha demostrado su utilidad en el diagnóstico precoz con mejoría en nutrición, función pulmonar y cognición. Se coordinaron dos centros para la medición de los valores de TIR y diagnóstico de fibrosis quística, usando un algoritmo tradicional y uno adaptado para mejorar resultados previos sin coordinación. Método. Se presentan datos del período pre-coordinación donde no hubo diagnósticos de FQ a partir de TIR. Se detallan datos del período coordinado, que aun no siendo cronológica ni numéricamente igual al anterior, muestran resultados totalmente distintos. Para TIR se utilizó DELFIA y para test de sudor iontoforesis por pilocarpina. Resultados. Período pre-coordinación entre centros (enero de 2005 a febrero de 2010): se realizaron 18.732 determinaciones de TIR. 74 fueron TIR (+). No hubo diagnósticos de FQ por problemas de seguimiento. Período coordinado: desde el 1° de marzo de 2010 hasta el 31 de diciembre de 2011. Se realizaron 41.142 determinaciones de TIR. Se encontró TIR elevado en la 1ª muestra en 91 niños (0,22%). Se diagnosticaron 4 fibroquísticos por medio de los algoritmos presentados. Conclusiones. En 71 RN seguidos efectivamente con TIR positivo, se diagnosticaron 4 FQ en el período coordinado. Disminuyó la edad media al diagnóstico de 4,54 años a 2,5 meses (aún pocos casos) y la cantidad de perdidos al seguimiento bajó de 36,95 % a 15,90%. Estas cifras sugieren un resultado positivo producto de la integración y centralización de los sistemas como lo cita toda la literatura.

Introduction. Neonatal screening in cystic fibrosis has shown to be an important tool for early diagnosis, better nutrition, diminished lung function and cognitive development. Our laboratory and CF Center were coordinated for IRT screening and diagnosis. We used a standard and a modified algorithm to improve previous results of an uncoordinated period. Method. IRT-IRT-sweat test algorithms was implemented using Dissociation-Enhanced Lanthanide Fluroimmunoassay (DELFIA) for IRT, and pilocarpine iontophoresis for sweat test. Pre-coordination period data are also presented (January 2005 - February 2010) when follow up was difficult .Details for the coordinated period are presented although no statistical comparisons are intended since the number of patients and time-periods studied were completely different. Results. a) pre-coordination period: 18,732 IRT measurements were done. 74 positive IRT tests were detected. No CF diagnosis was made by IRT screening due to problems with follow-up. Only 3 CF patients were diagnosed by genetic studies requested only due to clinical symptoms. b) Coordinated period (March 2010 to December 2011): 41,142 IRT samples were analized. 91 positive first sample IRT detected (0.22%). 71 patients remained for effective follow-up. Four CF diagnosis were made by the algorithm IRT screening-sweat test. Conclusions. In 71 remaining patients after first IRT positive result, CF was diagnosed in four patients. Mean age of diagnosis diminished from 4.54 years to 2.5 months although numbers are small for definitive conclusions. These figures suggest that positive results are the result of coordination and centralization in the CF Center as suggested by many references.

Research progress of autophagy and its effect on acute pancreatitis / 国际外科学杂志

Acute pancreatitis (AP) is activated trypsin-induced pancreatic and its peripheral tissue inflammation caused by their own digestion.The activation in advance of trypsinogen and the inflammation cascade in pancreatic acinar cells are thought to be a key mechanism of the onset and development of AP.Autophagy pathway acting as Ⅱ type of programmed cell death occurs in the early pathological course of AP,blockade of which contributes to aggravating necrosis of acinar cells in AP.This article mainly discussed the recent advances in the understanding of autophagy researches and its function in the mechanism of AP.

Changes and its significance of autophagy in rats with acute necrosis pancreatitis / 中华消化杂志

Objective To investigate the changes and significance of autophagy in rats with experimental acute necrosis pancreatitis (ANP).Methods According to method of random number,18 rats were randomly divided into control group,ANP group,ANP+rapamycin (RAP) group.The ANP rat model was established by intraperitoneal injection of 20％ L-arginine.The rats of ANP+RAP group were intraperitoneal injected with RAP 1.2 mg/kg at 30 minutes before modeling.The rats of control group were intraperitoneal injected with 0.9％ NaCl solution.The blood was drawed from the hearts nine hours after modeling for subsequent experiments.Serum levels of trypsinogen activation peptide (TAP),interleukin (IL-1),IL-6 and tumor necrosis factor (TNF) α were measured with enzyme-linked immunosorbent assay.The pancreatic tissues were pathologically scored.Autophagy-related structures in rat pancreatic acinar cells were observed by transmition electron microscopy.The expression of autophagy marker microtuble assciated protein 1 light chain 3 (LC3)-Ⅱ and Beclin-1 at mRNA and protein level were measured by quantitative real-time polymerase chain reaction (qRT-PCR),Western bloting and immunohistochemistry.The single factor analysis of variance was used for mean comparison among groups.Results A rat model of ANP was successfully established.Histopathological score of pancreas acinar cell necrosis of ANP+RAP group (2.19±1.38) was higher than that of ANP group (0.97±0.68),and the difference was statistically significant(F=33.75,P＜0.05).The results of Western blotting indicated that the protein expression of LC3-Ⅱ and Beclin-1 in ANP group (35.25±2.68 and 49.40±5.28)were higher than those in control group (1.54±0.16 and 0.78±0.06),furthermore the expressions in ANP+RAP group(123.53±3.21 and 76.41±3.80) were higher than those in ANP group,and the differences were statistically significant(F=2 045.54,326.87,both P＜0.01).Immunohistochemistry results also indicated that the LC3Ⅱ and Beclin-1 expression at protein level of ANP+RAP group (7 570.63±4 357.67 and 3 418.09±2 035.78) were higher than those of ANP group (1 926.53±1 414.44 and 536.11±403.10),and the differences were statistically significant (F=39.83,41.58,both P＜0.01).The expression of Beclin-1 at mRNA level of ANP group (107.12±29.10) was statistically higher than that of control group(7.01 ±3.39),and the difference was statistically significant (F=3.61,P＜0.05),but the expression of ANP+RAP group (97.63 ± 65.38)was no significant difference compared with ANP group.However,the expression of LC3-Ⅱ at mRNA level of ANP+ RAP group (4.37 ± 1.67) was statistically higher than that of ANP group (1.76 ± 1.59),and the difference was statistically significant(F=16.10,P＜0.05),but the expression of ANP group was no significant difference compared with control group (1.51 ±0.95).The result of electron microscopy showed that autophagy related structures increased in ANP group compared with that of control group,which of ANP+RAP group was more.The serum levels of TAP,IL-1 and IL-6 of ANP + RAP group were (36.47 ± 1.71) pmol/L,(122.88± 26.67) pg/mL and (107.39±13.95) pg/mL,which were all higher than those of ANP group ((25.63 ± 6.05) pmol/L,(98.06 ±9.29) pg/mL and (86.16± 7.20) pg/mL),and the differences were statistically significant (F=116.71,50.45,79.67; all P＜0.01).There was no significant difference in TNFα between ANP+ RAP group ((140.80±60.82) pg/mL) and ANP group ((105.23±6.95) pg/mL,F=14.76,P＞0.05).Conclusions Autophagy increased in rats with ANP.Promoting autophagy could significantly activate trypsinogen,aggravate pancreatic injury and increase inflammation reaction,which indicated that autophagy might involve in the pathogenesis of ANP through trypsinogen activation.

The role of trypsinogen activation peptide in the early diagnosis of severe acute pancreatitis and the prediction of outcome / 临床外科杂志

Objective To explore the role of trypsinogen activation peptide( TAP)in the early di-agnosis of severe acute pancreatitis( AP)and the prediction of outcome. Methods Eighty-nine cases with pancreatitis were collected containing 45 cases of severe AP and 44 of mild AP. 32 cases of non-AP acute abdominal disease were chosen as control group. Blood of all patients was collected within 6h to measure TAP. ROC curve was used to analyze the results. Results The TAP was different among the three groups (p<0. 01);the TAP of patients with severe AP was the highest. ROC curve indicated that 2. 78 nmol/L was the best cutoff to define AP and non-AP;the sensitivity,specificity and accuracy was 88. 8%,100%and 91. 5%,respectively. 8. 55 nmol/L was used to define severe AP and mild AP;the sensitivity,speci-ficity and accuracy was 92. 9%,95. 2% and 88. 7%,respectively. 11. 20 nmol/L was used to predict the occurrence of complications;the sensitivity,specificity and accuracy was 75. 0%,90. 4% and 82. 2%,re-spectively. Conclusion The TAP can be used to diagnose AP,especially to identify severe AP. The TAP is also beneficial for predicting the outcome of patients with severe AP.

Role of autophagy in development and progression of acute pancreatitis / 临床肝胆病杂志

Acute pancreatitis is considered an autodigestive disorder in which inappropriate activation of trypsinogen to trypsin within pancre-atic acinar cells leads to the development of pancreatitis.Autophagy is an evolutionarily preserved degradation process of cytoplasmic cellular constituents,and it is one of the early pathological processes in acute pancreatitis.Autophagic flux is impaired in acute pancreatitis,which mediates the key pathologic responses of this disease.Impaired autophagy,dysfunction of lysosomes,and dysregulation of autophagy suggest a disorder of the endolysosomal pathway in acute pancreatitis.The role of autophagy in acute pancreatitis is discussed from the aspects of au-tophagic process,autophagy and activation of trypsinogen,impaired autophagy and acute pancreatitis,and defective autophagy promoting in-flammation.

Changes of zymophagy during cell model of caerulein-induced experimental acute pancreatitis / 中华消化杂志

Objective To observe the changes of zymophagy during experimental acute pancreatitis (AP) induced by caerulein.Methods Pancreatic acinar cell line AR42J cells were cultured in 6-well plates till 90％ confluent and then divided into AP group and control group.Caerulein (1 × 10-8 mol/L) was added into AP group to establish AP cell model,and 1640 cell culture medium was added into control group.After caerulein treated for one,four,six,eight,12 and 24 hours,cells and cell culture supernatant were collected.The levels of cytokine interleukin (IL)-1,tumor necrosis factor (TNF)α,trypsinogen activation (TAP) and amylase were measured with enzyme-linked immunosorbent assay (ELISA) method.The expression of LC3 and Beclin1 at mRNA of each group were detected by reverse transcription-polymerase chain reaction (RT-PCR).The LC3B protein level of each group were detected by Western blotting.The changes of autophagosome and zymophagosome were observed by transmission electron microscopy.The difference between AP group and control group was analyzed by analysis of variance.Results The level of IL-1,TNFα,amylase and TAP in cell culture supernatant of control group was (18.83±7.10) pg/mL,(14.20±3.79) pg/mL,(10.03±2.85) U/L and (39.48±8.62) pg/mL,respectively.Those of AP group significantly increased at first hour ((62.13±11.25) pg/mL,F=3.32,P＜0.01 ; (30.98±7.11) pg/mL,F=3.05,P＜0.05; (25.06±6.82) U/L,F=2.90,P＜0.05 and (128.51± 18.30) pg/mL),F=2.62,P＜0.01,at fourth or sixth hour reached peak (IL-1 at fourth hour:(71.96± 15.82) pg/mL,F=7.25,P＜0.01;TNFα at sixth hour:(39.92±8.94) pg/mL,F=4.93,P＜0.05; amylase at fourth hour:(28.83 ± 8.31) U/L,F=2.06,P＜0.05; TAP at fourth hour:(146.29± 29.36) pg/mL,F=0.14,P＜0.01) and then gradually decreased.At fourth and sixth hour,the expression of LC3 at mRNA level in AP group was 3.18±0.82,1.71±0.14,respectively,while the expression of Beclin-1 rnRNA at first,fourth hour was 2.44±0.34 and 4.13±0.30,all of them were significantly increased compared with those of control group (0.21±0.04 and 0.30±0.08,LC3 mRNA F=0.79、0.06; Beclin mRNA F=2.31、0.36,all P＜ 0.05).There were no significant differences at other time points.The numbers of autophagosome and zymophagosome of AP group were significantly higher than those of control group under transmission electron microscopy.Conclusion Zymophagy occurred during AP cell model induced by caerulein,which suggested that zymophagy might involve in the mechanism of AP.

The relation of cationic trypsinogen gene G191R mutation and pathogenesis of pancreatitis / 中华胰腺病杂志

Objective To observe the prevalence of anionic trypsinogen (PRSS2) gene G191R mutation in patients with acute pancreatitis (AP) and chronic pancreatitis (CP),and to investigate the effect of PRSS2 gene G191R mutation on susceptibility to pancreatitis.Methods The blood samples of 82 patients with acute pancreatitis,73 patients with chronic pancreatitis and 138 healthy subjects were collected,and genomic DNA was extracted.Nest PCR were performed to amplify PRSS2 gene and restriction fragment length polymorphism (RFLP) was followed by using Hpy188Ⅲ to distinguish the G191R mutation.DNA sequencing analysis was performed to confirm the mutation status.Results The size of nest PCR products was 436 bp.RFLP2 produced 309 bp and 127 bp fragments,which were resulted from PRSS2 gene G191R mutation (GGA →AGA).DNA sequencing analysis of the PCR products further confirmed the PRSS2 gene G191R mutation.Five of eighty-two(6.1％) patients with acute pancreatitis had PRSS2 gene G191R mutation (OR=0.682,95％ CI 0.231 ～ 2.010); one of seventy-three (1.4％) patients with chronic pancreatitis had the mutation (OR =0.145,95％ CI 0.019 ～ 1.145),and the corresponding value in healthy group was 8.7％ (12/138).The G191R mutation rate in patients with chronic pancreatitis was significantly lower than that in healthy group (x2 =0.432,P =0.035),but the G191R mutation rates were not significantly different between AP group and healthy group (x2 =0.487,P =0.485).Conclusions PRSS2 gene G191R mutation facilitates the degradation of anionic trypsin,and may reduce the incidence of chronic pancreatitis.

Neonatal screening for cystic fibrosis in São Paulo State, Brazil: a pilot study

Cystic fibrosis is one of the most common autosomal recessive hereditary diseases in the Caucasian population, with an incidence of 1:2000 to 1:3500 liveborns. More than 1000 mutations have been described with the most common being F508del. It has a prevalence of 23-55 percent within the Brazilian population. The lack of population-based studies evaluating the incidence of cystic fibrosis in São Paulo State, Brazil, and an analysis concerning the costs of implantation of a screening program motivated the present study. A total of 60,000 dried blood samples from Guthrie cards obtained from April 2005 to January 2006 for neonatal screening at 4 reference centers in São Paulo State were analyzed. The immunoreactive trypsinogen (IRT)/IRT protocol was used with the cut-off value being 70 ng/mL. A total of 532 children (0.9 percent) showed IRT >70 ng/mL and a 2nd sample was collected from 418 (80.3 percent) of these patients. Four affected children were detected at two centers, corresponding to an incidence of 1:8403. The average age at diagnosis was 69 days, and 3 of the children already showed severe symptoms of the disease. The rate of false-positive results was 95.2 percent and the positive predictive value for the test was 8 percent. The cost of detecting an affected subject was approximately US$8,000.00 when this cystic fibrosis program was added to an existing neonatal screening program. The present study clearly shows the difficulties involved in cystic fibrosis screening using the IRT/IRT protocol, particularly in a population with no long-term tradition of neonatal screening.

Clinical evaluation of rapid urinary trypsinogen-2 assay in the detecting and screening of acute pancreatitis / 中华胰腺病杂志

the specificity was 95%.Conclusions Urinary trypsinogen-2 assay test strip was rapid,convenient and sensitive for the diagnosis of AP with low cost,which was worth applying in emergency department for patients with acute abdomen.

The value of serum and urine trypsinogen activation peptide in the diagnosis and severity predicting of acute pancreatitis / 中国医师进修杂志

Objective To explore the value of trypsinogen activation pepfide (TAP) levels in blood plasma and urine in the diagnosis of acute pancreafitis at early stage and severity predicting combined with the CT results. Method Sixty-five patients with acute pancreatitis who scanned with enhanced CT were di-vided into mild acute pancreatitis (MAP) group and severe acute pancreatitis (SAP) group according to the CT results, 29 patients with acute abdomen except for acute pancreatitis were as control group. TAP levels in blood plasma and urine were measured in all patients. Results At the time of 6, 12 and 24 h after admis-sion, the level of blood plasma TAP in SAP group ( > 9.0 nmol/L) was significantly higher than that in MAP group(< 3.5 nmol/L), there were significant differences between SAP group and MAP group or control group (P<0.05). And the median urinary TAP in SAP group (102.8, 78.2, 52.3 nmol/L)was significantly higher than those in MAP group (32.5, 28.7, 25.6 nmol/L)and control group(12.6, 12.2, 11.5 nmol/L), there were significant differences among three groups (P<0.05). At the time of 3, 5 d after admission, there was no significant difference in TAP levels in blood plasma and urine among three groups (P>0.05). Conclusions Early dynamic detection of TAP levels in blood plasma and urine is of higher diagnostic value and predictive value for acute pancreatitis, which is of benefit in the early diagnosis and treatment of acute pancreatitis.

Relationship between Carbachol Hyperstimulation-induced Pancreatic Intracelluar Trypsinogen and NF-кB Activation in Rats in vitro / 华中科技大学学报（医学）（英德文版）

The relationship between intracelluar trypsinogen activation and NF-r,B activation in rat pancreatic acinar cells induced by M3 cholinergic receptor agonist (carbachoi) hyperstimulation was studied. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, the active pro- tease inhibitor (pefabloc) and NF-кB inhibitor (PDTC) in vitro. Intracelluar trypsin activity was measured by using a fluorogenie substrate. The activity of NF-кB was monitored by using electro- phoretic mobility shift assay. The results showed that after pretreatment with 2 mmol/L pefabloc, the activities of trypsin and NF-кB 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 addi- tion of 10-2mol/L PDTC resulted in a significant decrease of NF-кB 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 trypsi- nogen activation is likely involved in the regulation of high concertrations of carbachol-induced NF-кB activation in pancreatic acinar cells in vitro. NF-кB activation is likely not necessary for high concertrations of carbachol-induced trypsinogen activation in pancreatic acinar cells in vitro.

Three Cases of Hereditary Pancreatitis in Two Households in the Same Family Associated with R122H Mutation in Cationic Trypsinogen Gene / 대한소화기학회지

Hereditary pancreatitis is a rare, autosomal dominant, inherited disease characterized by recurrent attacks of acute pancreatitis with the development of chronic pancreatitis and an increased risk of pancreatic cancer. R122H or N29I mutation in cationic trypsinogen (protease serine 1, PRSS1) gene causes hereditary pancreatitis. R122H mutation is the most common mutation that causes pancreatitis by preventing deactivation of trypsin within the pancreas and prolonging its action. Three members of the family, the patient, her elder son, and her niece experienced recurrent attacks of pancreatitis. We analyzed five exons of the PRSS1 gene in DNA samples of five family members including her husband and younger son who were asymptomatic. We found out that four members of the family, the patient, her two sons, and her niece, had R122H mutation in the exon 3 of PRSS1 gene. Finally, we diagnosed hereditary pancreatitis in two households in the same family.

Establishment and evaluation of indirect competitive enzyme-linked immunosorbent assay for detecting the concentration of trypsiongen activation peptides / 国际检验医学杂志

Objective To establish an ELISA method for detection of the concentration of TAP in human urine samples.Methods TAP-BSA was used as coating antigen. The TAP was a competitor to TAP-BSA. They reacted to the limited amount of monoclonal antibody against TAP.Results The optimal concentration of the coating antigen was 250ng/ml.The dilutions of monolclonal antibody against TAP and sheep anti-mouse IgG were 25?g/ml and 1∶4 000 respectively. The optimal range was from 0.69 to 1 000ng/ml.The assay provided a sensitivity of 0.69ng/ml.The coefficients of variation of intra-assay and inter-assay were 9.10% and 10.33% respectively. The average recovery rate of TAP was 97.70%.Conclusion The indirect competitive enzyme-linked immunosorbent assay is established.

Relationship between Carbachol Hyperstimulation-Induced Pancreatic Acinar Cellular Injury and Trypsinogen or NF-κB Activation in Rats in vitro / 华中科技大学学报（医学）（英德文版）

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 cells 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 cells. 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 10-2 mol/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.

A case of hereditary pancreatitis with a N29I mutation in the cationic trypsinogen gene / 소아과

Hereditary pancreatitis is an autosomal dominant disease characterized by recurrent episodes of pancreatitis, often beginning in childhood, with a positive family history involving at least two other affected family members with no known other precipitating factors. Most forms of hereditary pancreatitis are caused by one of two common mutations, i.e., R122H in exon 3 and N29I in exon 2 of the cationic trypsinogen (CT) (PRSS1) gene, located on chromosome 7. The authors describe the case of a 15-year-old boy who had suffered from recurrent attacks of pancreatitis since age three. His mother and grandmother had chronic pancreatitis and diabetes mellitus. Mutation analysis was performed on the family due to the suspicion of hereditary pancreatitis. The CT gene was analyzed in DNA samples extracted from the peripheral blood of three family members, the mother, the proband, and the proband's sister. Two members of the family, the mother and the proband, were found to have a N29I mutation in the CT gene. The authors document the first family with hereditary pancreatitis associated with the N29I mutation in Korea.