Energy values and protein digestibility of soybean milk by-product in pigs based on in vitro assays / Valores energéticos y digestibilidad proteica del subproducto de la leche de soja en cerdos con base en ensayos in vitro / Valores energéticos e digestibilidade protéica do subproduto do leite de soja em suínos com base em ensaios in vitro

Abstract Background: Soybean milk by-product (SMBP) is a potential alternative feed ingredient in swine diets due to its high protein content. However, information on energy and nutritional values of SMBP used as swine feed ingredient is limited. Objective: To estimate energy values and protein digestibility of SMBP in pigs based on in vitro assays. Methods: Four SMBP samples were obtained from 3 soybean milk-producing facilities. In vitro total tract disappearance (IVTTD) and in vitro ileal disappearance (IVID) of dry matter (DM) in the SMBP samples were determined. In vitro ileal disappearance of crude protein was determined by analyzing crude protein content in undigested residues after determining IVID of DM. Digestible and metabolizable energy of SMBP were estimated using gross energy, IVTTD of DM, and prediction equations. Results: Sample 4 had greater IVTTD of DM than that of sample 3 (97.7 vs. 94.4%, p<0.05), whereas IVID of DM in sample 4 was lower compared with sample 1 (53.5 vs. 65.0%, p<0.05). In vitro ileal disappearance of crude protein in sample 2 was greater than that in sample 1 and 3 (92.6 vs. 90.6 and 90.1%; p<0.05). The estimated metabolizable energy of SMBP ranged from 4,311 to 4,619 kcal/kg as-is basis and the value of sample 3 was the least (p<0.05) among SMBP samples. Conclusion: Energy values and protein digestibility should be determined before using SMBP in swine diets.

Resumen Antecedentes: El subproducto de la leche de soja (SMBP) es un ingrediente alimenticio alternativo con uso potencial en dietas porcinas dado su alto contenido de proteína. Sin embargo, la información sobre sus valores energéticos y nutricionales para alimentación de cerdos es muy limitada. Objetivo: Estimar los valores de energía y la digestibilidad de la proteína del SMBP en cerdos con base en ensayos in vitro. Métodos: Se obtuvieron cuatro muestras de SMBP de tres empresas productoras de leche de soja. Se determinaron la desaparición de tracto total in vitro (IVTTD) y la desaparición ileal in vitro (IVID) de la materia seca (DM) en las muestras de SMBP. La desaparición ileal in vitro de proteína cruda se determinó analizando el contenido de proteína cruda en residuos no digeridos después de determinar la IVID de la DM. La energía digestible y metabolizable de SMBP se estimó utilizando la energía bruta, IVTTD de la DM y ecuaciones de predicción. Resultados: La muestra 4 tuvo una mayor IVTTD de la DM que la muestra 3 (97,7 vs. 94,4%, p<0,05), mientras que la IVID de la DM en la muestra 4 fue menor en comparación con la muestra 1 (53,5 vs. 65,0%, p<0,05). La desaparición ileal in vitro de la proteína cruda en la muestra 2 fue mayor que la de las muestras 1 y 3 (92,6 vs. 90,6 y 90,1%; p<0,05). La energía metabolizable estimada de SMBP varió de 4.311 a 4.619 kcal/kg (en base húmeda) y el valor de la muestra 3 fue el menor (p<0.05) entre las muestras de SMBP. Conclusión: Los valores de energía y la digestibilidad de la proteína deben determinarse antes de usar el SMBP en dietas porcinas.

Resumo Antecedentes: O subproduto do leite de soja (SMBP) é um potencial ingrediente alternativo na dieta de suínos, considerando seu alto teor de proteínas. No entanto, as informações sobre os valores energéticos e nutricionais do SMBP usado como ingrediente alimentar para suínos são limitadas. Objetivo: Estimar valores energéticos e digestibilidade protéica do SMBP em suínos com base em ensaios in vitro. Métodos: Foram obtidas quatro amostras de SMBP de três instalações produtores de leite de soja. Foram determinados o desaparecimento total do trato in vitro (IVTTD) e o desaparecimento ileal in vitro (IVID) da matéria seca (DM) nas amostras de SMBP. O desaparecimento ileal in vitro da proteína bruta foi determinado pela análise do conteúdo de proteína bruta em resíduos não digeridos após a determinação da IVID do DM. A energia digerível e metabolizável do SMBP foi estimada usando energia bruta, IVTTD do DM e equações de predição. Resultados: a amostra 4 apresentou maior IVTTD de DM do que a amostra 3 (97,7 vs. 94,4%, p<0,05) enquanto a IVID do DM na amostra 4 foi menor em comparação com a amostra 1 (53,5 vs. 65,0%, p<0,05). O desaparecimento ileal in vitro da proteína bruta na amostra 2 foi superior ao da amostra 1 e 3 (92,6 vs. 90,6 e 90,1%; p<0,05). A energia metabolizável estimada do SMBP variou de 4.311 a 4.619 kcal/kg no estado em que se encontra e o valor da amostra 3 foi o menor (p<0,05) entre as amostras do SMBP. Conclusão: os valores energéticos e a digestibilidade das proteínas devem ser determinados antes do uso do SMBP nas dietas suínas.

Relationship between mechanism of ulinastatin reducing perioperative myocardial injury and ferroptosis in peripheral blood mononuclear cells in pediatric patients undergoing heart surgery under cardiopulmonary bypass / 中华麻醉学杂志

Objective:To investigate the relationship between the mechanism of ulinastatin reducing perioperative myocardial injury and ferroptosis in peripheral blood mononuclear cells (PBMCs) in pediatric patients undergoing heart surgery under cardiopulmonary bypass (CPB).Methods:A total of 60 pediatric patients of either sex, aged 4-8 yr, of American Association of Anesthesiologists physical status Ⅱ or Ⅲ, undergoing elective repair of ventricular septal defect under CPB, were divided into 2 groups by a random number table method: control group (C group) and ulinastatin group (UTI group), with 30 cases in each group.Combined intravenous-inhalational anesthesia was used.In UTI group, ulinastatin 20 000 U/kg was diluted to 100 ml in normal saline, 50 ml was infused through the central vein over 15 min starting from 20 min before skin incision, and the remaining 50 ml was instilled through the CPB pipeline over 15 min starting from 10 min of CPB.The equal volume of normal saline was given instead in C group.Blood samples from the internal jugular vein were collected after anesthesia induction and before skin incision (T 1), at 30 min after start of CPB (T 2), immediately after termination of CPB (T 3) and at 24 h after termination of CPB (T 4) for determination of the levels of amino-terminal B-type pro-brain natriuretic peptide (NT-proBNP), cardiac troponin I (cTnI) and creatine kinase isoenzymes (CK-MB) in plasma by enzyme-linked immunosorbent assay.PBMCs were extracted by modified Ficoll density gradient centrifugation method for determination of the concentrations of Fe 2+ and malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in PBMCs (by colorimetric method) and expression of long-chain acyl-CoA synthase 4 (ACSL4) and glutathione peroxidase 4 (GPX4) in PBMCs (by Western blot). Results:Compared with the baseline at T 1, the levels of NT-proBNP, cTnI and CK-MB in plasma were significantly increased, the concentrations of Fe 2+ and MDA in PBMCs were increased, the expression of ACSL4 in PBMCs was up-regulated, and the activity of SOD was decreased, and the expression of GPX4 was down-regulated at T 2-4 in two groups ( P<0.05). Compared with C group, the plasma levels of NT-proBNP, cTnI and CK-MB were significantly decreased, the concentrations of Fe 2+ and MDA in PBMCs were decreased, the expression of ACSL4 in PBMCs was down-regulated, the activity of SOD was increased, and the expression of GPX4 was up-regulated at T 2-4 in UTI group ( P<0.05). Conclusion:The mechanism by which ulinastatin reduces perioperative myocardial injury may be related to inhibition of ferroptosis in PBMCs in the pediatric patients undergoing open heart surgery under CPB.

Effect of ulinastatin on expression of metabotropic glutamate receptor Ⅱ during cerebral ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To evaluate the effect of ulinastatin on the expression of metabotropic glutamate receptor Ⅱ ( mGluRⅡ) during cerebral ischemia-reperfusion ( I∕R) in rats. Methods Forty-eight male Sprague-Dawley rats, aged 6-8 weeks, weighing 230-280 g, were divided into 3 groups ( n=16 each) by a random number table method: sham operation group (S group), cerebral I∕R group (I∕R group) and ulinastatin group ( U group) . The model of cerebral I∕R injury was established by occluding the right middle cerebral artery using a nylon thread with a rounded tip inserted into internal carotid artery and advanced cranially until resistance was met. Middle cerebral artery occlusion was maintained for 2 h followed by 24-h reperfusion. Ulinastatin 100000 U∕kg was injected via the tail vein immediately after onset of reperfusion in group U. The neurological deficit score ( NDS) was assessed after 24 h of reperfusion. The rats were then sacrificed, and brain tissues were obtained for determination of brain infarction ( by TTC staining) , expression of IκB-α in cerebral ischemic penumbra ( by Western blot) and expression of mGluRⅡ in cerebral ischemic penumbra ( by immunofluorescent staining) . The percentage of cerebral infarct vol-ume was calculated. Results Compared with S group, the NDS and percentage of cerebral infarct volume were significantly increased, the expression of mGluRⅡ in cerebral ischemic penumbra was up-regulated, and the expression of IκB-α in cerebral ischemic penumbra was down-regulated in I∕R and U groups ( P<0. 05). Compared with I∕R group, the NDS and percentage of cerebral infarct volume were significantly de-creased, the expression of mGluRⅡ in cerebral ischemic penumbra was down-regulated, and the expres-sion of IκB-α in cerebral ischemic penumbra was up-regulated in U group ( P<0. 05) . Conclusion The mechanism by which ulinastatin mitigates cerebral I∕R injury may be related to inhibiting the expression of mGluR Ⅱ in cerebral ischemic penumbra of rats.

Effect of Edaravone Combining Ulinastatin on Brain Protection in Patients of Type A Aortic Dissection After Total Arch Replacement / 中国循环杂志

Objective: To observe the effect of edaravone combining ulinastatin on brain protection in patients of type A aortic dissection (AAD) after total arch replacement. Methods: A total of 60 AAD patients with total arch replacement in our hospital from 2014-09 to 2016-01 were prospectively studied. Based on peri-operative application of edaravone and ulinastatin, the patients were divided into 2 groups: EU group: 1) the patients received ulinastatin 300000 U/8h and edaravone 0.5mg/Kg/12h from administration to 3 days post-operation, 2) during cardiopulmonary bypass, the patients received ulinastatin 300000 U/2h and edaravone 0.5mg/Kg; Control group, the patients had no such treatment.n=30 in each group. The following items were observed:①operative condition;②blood levels of speciifc brain injury markers as S-100 and neuron speciifc enolase (NSE) at different time points: beginning of surgery (T0), opening aorta clamp (T1), right after cardiopulmonary bypass (T2), entering ICU (T3), 24h post-operation (T4) and 3 days post-operation (T5); ③post-operative condition. Results:①Durations of operation, cardiopulmonary bypass, cardiac arrest and bilateral antegrade selective cerebral perfusion (BACP), the frequency of BACP and UACP (unilateral antegrade selective cerebral perfusion), the lowest rectal temperature and blood levels of S-100, NSE at T0 were similar between 2 groups.②Compared with Control group, EU group had decreased S-100 and NSE from T1 to T5,P0.05. Conclusion: Edaravone combining ulinastatin had brain protective effect in AAD patients after total arch replacement;it may reduce blood speciifc brain injury markers while the clinical signiifcance should be further investigated.

A novel thermosensitive in-situ gel of gabexate mesilate for treatment of traumatic pancreatitis: An experimental study / 华中科技大学学报（医学）（英德文版）

Gabexate mesilate (GM) is a trypsin inhibitor, and mainly used for treatment of various acute pancreatitis, including traumatic pancreatitis (TP), edematous pancreatitis, and acute necrotizing pancreatitis. However, due to the characteristics of pharmacokinetics, the clinical application of GM still needs frequently intravenous administration to keep the blood drug concentration, which is difficult to manage. Specially, when the blood supply of pancreas is directly damaged, intravenous administration is difficult to exert the optimum therapy effect. To address it, a novel thermosensitive in-situ gel of gabexate mesilate (GMTI) was developed, and the optimum formulation of GMTI containing 20.6% (w/w) P-407 and 5.79% (w/w) P188 with different concentrations of GM was used as a gelling solvent. The effective drug concentration on trypsin inhibition was examined after treatment with different concentrations of GMTI in vitro, and GM served as a positive control. The security of GMTI was evaluated by hematoxylin-eosin (HE) staining, and its curative effect on grade II pancreas injury was also evaluated by testing amylase (AMS), C-reactive protein (CRP) and trypsinogen activation peptide (TAP), and pathological analysis of the pancreas. The trypsin activity was slightly inhibited at 1.0 and 5.0 mg/mL in GM group and GMTI group, respectively (P<0.05 vs. P-407), and completely inhibited at 10.0 and 20.0 mg/mL (P<0.01 vs. P-407). After local injection of 10 mg/mL GMTI to rat leg muscular tissue, muscle fiber texture was normal, and there were no obvious red blood cells and infiltration of inflammatory cells. Furthermore, the expression of AMS, CRP and TAP was significantly increased in TP group as compared with control group (P<0.01), and significantly decreased in GM group as compared with TP group (P<0.01), and also slightly inhibited after 1.0 and 5.0 mg/mL GMTI treatment as compared with TP group (P<0.05), and significantly inhibited after 10.0 and 20.0 mg/mL GMTI treatment as compared with TP group (P<0.01). HE staining results demonstrated that pancreas cells were uniformly distributed in control group, and they were loosely arranged, partially dissolved, with deeply stained nuclei in TP group. Expectedly, after gradient GMTI treatment, pancreas cells were gradually restored to tight distribution, with slightly stained nuclei. This preliminary study indicated that GMTI could effectively inhibit pancreatic enzymes, and alleviate the severity of trauma-induced pancreatitis, and had a potential drug developing and clinic application value.

Microproteínas urinarias en diabéticos tipo 2 normoalbuminúricos / Urinary microproteins in normoalbuminuric type 2 diabetic patients / Microproteínas urinárias em diabéticos tipo 2 normoalbuminúricos

En pacientes diabéticos tipo 2 normoalbuminúricos se observa la presencia de Microproteínas Urinarias (MU) en el rango 68-25 kDa. El objetivo del trabajo fue identificar en distintos estadios de la nefropatía diabética si dicho rango corresponde a un marcador de daño tubular. Se estudiaron 119 orinas espontáneas de pacientes diabéticos tipo 2; se les midió la relación albúmina/creatinina urinaria y la creatinina sérica. Se dividieron en 5 grupos: 71 normoalbuminúricos, 28 microalbuminúricos, 12 macroalbuminúricos, 2 urémicos en pre-diálisis y 6 en hemodiálisis. Las MU se detectaron en geles de poliacrilamida en 2 dimensiones para uso clínico y se analizaron con el programa Image J 1.30v. La identificación de las MU se realizó por “immunoblotting” o por espectrometría de masa MALDI-TOF-TOF. El 66% de los normoalbuminúricos presentaron las siguientes MU: orosomucoide, fragmento de 35 kDa de la cadena pesada H4 del inter alfa I inhibidor de tripsina y Beta Trace, las cuales no reflejaron daño tubular debido a que la concentración de las mismas no se incrementó en los pacientes en hemodiálisis, en comparación con los normoalbuminúricos. Dichas proteínas están vinculadas al endotelio vascular y podrían constituir un marcador urinario vascular-tubular renal de utilidad clínica en patologías sistémicas con riesgo cardiovascular y funcionalidad renal conservada.

Urinary excretion of microproteins (MU) was detected in normoalbuminuric type 2 diabetic patients, in the range of 68-25 kDa using SDS-PAGE with silver staining. The purpose of this study was to identify MU in diabetic patients in different grades of diabetic nephropathy, in order to clarify the diagnostic relevance as a marker of renal tubular damage. In the spontaneous urine of 119 type 2 diabetic patients, urinary albumin, urinary creatinine and serum were determined. Five groups were formed: 71 normoalbuminuric, 28 microalbuminuric, 12 macroalbuminuric, 2 in pre-dialysis and 6 in hemodialysis. The MU were separated by two-dimensional polyacrylamid gel electrophoresis for clinical use (2D UC) and were identified by immunoblotting or MALDI-TOF-TOF mass spectrometry and analyzed using Image J version 1.30v. Of the normoalbuminurics patients studied, 66% excreted the following MU: orosomucoid, 35 kDa fragment of inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4) and Beta Trace; but their concentrations did not reflect tubular damage because they exhibited a progressive downregulation. These proteins are involved in vascular endothelium, and they could be a marker of renal tubular-microvascular disease that would be useful in systemic diseases with cardiovascular risk and with preserved renal function.

Em pacientes diabéticos tipo 2 “normoalbuminúricos” se observa a presença de microproteínas urinárias (MU) em um intervalo compreendido entre 68 e 25 kDa. O objetivo do trabalho é identificar em distintos estágios da nefropatia diabética se tal intervalo corresponde a um marcador de dano tubular. Foram estudadas 119 amostras de urina espontânea de pacientes diabéticos tipo 2; foi medida a reação albumina/creatinina urinária e a creatinina sérica. Dividiram-se em 5 grupos: 71 normoalbuminúricos, 28 microalbuminúricos, 12 macroalbuminúricos, 2 urêmicos pré-dialise e 6 em hemodiálise. Foram detectadas as MU em géis de poliacrilamida em duas dimensões para o uso clínico e analisadas com o programa Image J versão 1.30v. A identificação das MU foi realizada por “immunoblotting” ou por espectrometria de massa MALDI-TOF-TOF. 66% de normoalbuminúricos apresentaram as seguintes MU: orosomucoide, Fragmento de 35 kDa da cadeia pesada H4 do interalfa inibidor da tripsina e Beta Trace, as quais não mostraram dano tubular devido a que a concentração das mesmas não está aumentada nos pacientes em hemodiálise, em comparação com os normoalbuminúricos. Estas proteínas estão envolvidas com o endotélio vascular e poderiam constituir um marcador urinário vascular-tubular renal de utilidade clínica em patologias sistêmicas com risco cardiovascular e funcionalidade renal conservada.

Construction of secretory expression vector of rhKD/APPvar and expression and purification of its recombinant protein in Pichia pastoris / 吉林大学学报(医学版)

Objective To construct the engineering bacteria expressing the recombinant human Kunitz protease inhibitor domain of amyloid protein precursor variant (rhKD/APPvar)in Pichia pastoris,and to establish the methods suitable for large-scale fermentation and purification of rhKD/APPvar.Methods The rhKD/APPvar expression vector was constructed based on the rhKD/APPvar-pPICZαexpression vector. Two restriction enzyme loci (ApaⅠ and SacⅡ)were added to two flanks of KD/APP and human KD/APP activity center RAM was replaced by the active site of BPTI KAR.After the rhKD/APPvar-pPICZαexpression vector was transformed into Pichiapastoris,optimized expression and purification of rhKD/APPvar was performed.The rhKD/APPvar was purified with cation exchange chromatography and desalting.Results The results of digestion identification and DNA sequencing analysis demonstrated that the recombinant plasmid rhKD/APPvar-pPICZα was successfully constructed and transfected into pastoris X-33. The SDS-PAGE analysis results indicated that rhKD/APPvar expressed after the induction of methanol and the relative molecular weight was 6 700.After a series of experiments the optimal expression conditions of rhKD/APPvar were obtained as follows:the optimal pH was 6.0 and the optimal induction time point was about the 5 th day for the strain.After purified the purity of rhKD/APPvar was about 95%.Conclusion KD/APPvar-pPICZ is successfully constructed;after expression in Pichia pastoris and purification,the rhKD/APPvar protein is obtained.

Deployment of gene specific marker in development of kunitz trypsin inhibitor free soybean genotypes.

Genetic elimination of kunitz trypsin inhibitor in soybean seed would obviate the need for boiling required to inactivate the antinutritional factor and therefore economize the soy processing. PI542044, the source of null variant of kunitz trypsin inhibitor gene is being used in the development of kunitz trypsin inhibitor free soybean genotypes in India. Gene specific marker can expedite the genetic elimination of this undesirable trait from popular soybean genotypes. In the present study, we tested the DNA amplification of soybean genotype PI542044 and kunitz trypsin inhibitor null lines derived from this genotype with a gene specific primer developed from the null variant of PI157740. The amplicons so obtained corresponded to the absence of kunitz trypsin inhibitor protein band on 10% polyacrylamide gel. The gene specific marker also amplified the null allele of template DNA of F1, BC1F1 and BC2F1 plants developed during marker assisted introgression of null allele of kunitz trypsin inhibitor into elite soybean cultivar JS97-52. The results presented show the utility of this gene specific marker developed from null allele of kunitz trypsin inhibitor for identification of kunitz trypsin inhibitor free genotypes developed from PI542044, the only source of null variant available in India.

The Effect of Urinary Trypsin Inhibitor Against Neuropathic Pain in Rat Models

BACKGROUND: Nerve injury sometimes leads to chronic neuropathic pain associated with neuroinflammation in the nervous system. In the case of chronic neuropathic pain, the inflammatory and algesic mediators become predominant and result in pain hypersensitivity following nervous system damage. It is well known that urinary trypsin inhibitor (ulinastatin, UTI) has an anti-inflammatory activity. Recently, the neuroprotective action of UTI on the nervous system after ischemic injury has been reported. Thus, we evaluated the neuroprotective effect of ulinastatin in a rat model of neuropathic pain. METHODS: Neuropathic pain was induced with L5 spinal nerve ligation (SNL) in male Sprague-Dawley rats weighing 100-120 g. The rats were divided into 3 groups, with n = 8 in each group. The rats in the control group (group 1) were administered normal saline and those in group 2 were administered UTI (50,000 U/kg) intravenously through the tail vein for 3 days from the day of SNL. Rats in group 3 were administered UTI (50,000 U/kg) intravenously from the 5th day after SNL. The paw withdrawal threshold was measured using the von Frey test for 3 days starting from the 5th day after SNL. RESULTS: The paw withdrawal thresholds were significantly increased in the rats of group 2 compared to the other groups (P < 0.05). CONCLUSIONS: Ulinastatin, which was administered for 3 days after SNL, increased the paw withdrawal threshold and it could have a neuroprotective effect in the rat model of neuropathic pain.

Evaluation of the shelf-life of vegetable-type soybean pods

The shelf-life of the vegetable-type soybean pods stored under different conditions was evaluated by chemical characteristics and color. The pods were harvested in the R6 stage and stored either at 30 or 7ºC for 9 d. After the storage period, the pods were blanched and threshed, and the immature green grains were used for the analysis. The protein content decreased after 6 d of storage at 7ºC. There was no difference in the lipid content after the storage at 30 and 7ºC for 9 d. The starch and sucrose contents decreased after the first day of storage at 30ºC. There was no difference in trypsin inhibitor activity until 6 d of storage at 30 and 7ºC. The green color of the pods that was an indication of the quality that was maintained when stored at 7ºC during 3 d. To preserve the quality of vegetable-type soybean, pods should be stored at 30ºC and consumed within 24 h or stored at 7ºC for up to 3 d of storage.

Effects of ulinastatin on myocardial injury in patients undergoing live donor liver transplantation / 中华麻醉学杂志

Objective To investigate the effects of ulinastatin on the myocardial injury in patients undergoing live donor liver transplantation.Methods Forty patients (AHA classification grade A or B),aged 40-64 yr,with a body mass index of 18-25 kg/m2,scheduled for live donor liver transplantation,were randomly divided into 2 groups ( n =20 each):control group (group C) and ulinastatin group (group U).Anesthesia was induced with midazolam,sufentanil,and cisatracurium besilate.The patients were tracheal intubated and mechanically ventilated.Ulinastatin 300 000 IU in 100 ml of normal saline was infused intravenously over 30 min after anesthesia induction and then the infusion was repeated at 4 h interval until the end of operation in group U,while the equal volume of normal saline was given in group C.Blood samples were taken from the central vein immediately before skin incision (T0,baseline),at 30 min of anhepatic phase (T1),at 30 min of neohepatic phase (T2),and at 0,4 and 24 h after operation (T3-5) for determination of the concentrations of serum cardiac troponin Ⅰ (cTnI),creatine kinase-MB (CK-MB) and N-terminal pro-brain natriuretic peptide (NT-proBNP).The changing rates of cTnI and CK-MB at T1-5 were calculated.The use of cardiovascular drugs and cardiovsscular accidents were recorded during operation.Results The serum cTnI,CK-MB and NT-proBNP concentrations were significantly higher at T2-5 than at T0 in the two groups ( P ＜ 0.05).Compared with group C,the serum cTnI,CK- MB and NT-proBNP concentrations at T2-5 were significantly deceased in group U ( P ＜ 0.05).The maximal changing rates of cTnI,CK-MB and NT-proBNP concentrations were 4.71 ± 1.62,6.85 ± 1.53 and 4.96 ± 1.23 respectively in group C,decreased to 3.26 ± 1.51,4.56 ± 1.62 and 3.67 ± 1.02 respectively in group U.There was no significant difference in the incidence of cardiovascular accidents and the use of dopamine between the two groups.Conclusion Intravenous infusion of ulinastatin can attenuate the myocardial injury to some extent in patients undergoing live donor liver transplantation.

Urinary trypsin inhibitor attenuates lipopolysaccharide-induced neutrophil activation / 대한마취과학회지

BACKGROUND: Urinary trypsin inhibitor (UTI), which is speculated to have anti-inflammatory effects, is one of serine protease inhibitors found in human urine and blood. The present study was conducted to clarify the effect of urinary trypsin inhibitor (UTI) on human neutrophil activation and its intracellular signaling mechanism in vitro. METHODS: To assess the possible interactions between UTI and lipopolysaccharide (LPS) in neutrophil activation, neutrophils from human blood were incubated with varying concentrations of UTI (1, 10, 100, 1,000 and 10,000 U/ml) plus LPS (100 ng/ml) or LPS alone in 24-well plates (5 x 106 cells/well). We measured protein levels for interleukin (IL)-6 and tumor necrosis factor-alpha (TNF-alpha) using enzyme-linked immunosorbent assay (ELISA) kits after 4 hours of incubation period. To elucidate the intracellular signaling pathway, we also measured the levels of phosphorylation of p38, ERK1/2 and JNK via Western blot analysis. Moreover, the nuclear levels of nuclear factor-kappa B (NF-kappaB) were determined with electrophoretic mobility shift assays (EMSA). RESULTS: UTI decreased the expression of inflammatory cytokines, including TNF-alpha and IL-6, and activation of intracellular signaling pathways, such as JNK, but not P38, ERK1/2 and nuclear translocation of NF-kappaB. CONCLUSIONS: UTI can attenuate LPS-induced neutrophil responses and may partially contribute to the treatment of neutrophil-mediated inflammatory diseases.

Improving islet yield and function by pancreatic ductal preservation / 中华肝胆外科杂志

Objective To determine whether pancreatic ductal preservation(pancreatic ductal injection of sparing TIUW solution at the time of pancreas procurement)can improve islet yield and function after cold preservation of pancreas.Methods Pancreases were classified into five groups:fresh pancreases(group1 ,n= 10);preserved for 6 hr in TIUW solution without and with pancreatic ductal preservation by TIUW solution(group2, n=10 and group3, n = 10);preserved for 24 hr in TIUW solution without and with pancreatic ductal preservation by TIUW solution(group4 ,n= 10 and group5,n= 10).Dithizon(DTZ)staining was used to identify islet morphous and yield, trypan blue uptake(TBU)was used to assess the nonviability of islets, the insulin secretory response to glucose was used to evaluate islet function in vitro, islet transplantation was used to determine islet function in vivo.Results Islet yields per pancreas after purification in group 1 to 5 were 590± 127, 272 ± 50,454 ± 65, 253± 56, 447 ± 66(islet equivalent ± SD), respectively.Percentage of nonviable islets in grouplto5were(5.7±4.2)%,(18.3±6.5)%,(11.7±4.2)%,(26.3±5.6)%,(15.0±5.3)%.Stimulation index in group 1 to 5 were 7.32±2.32, 4.81±1.17, 7.56±2.44, 2.88±1.00, 5.71±1.90.Cure rates in group 1 to 5 were 100%, 0%, 100%, 0%, 70%.In this study, the differences were significant between control groups and experimental groups(P＜0.05, group 2 vs.group3 and group4 vs.group5).Conclusion The pancreatic ductal preservation can improve islet yield and function.

Effect of ulinastatin on cytokine reaction during gastrectomy / 대한마취과학회지

BACKGROUND: Inflammation plays an important role in the postoperative morbidity of organs, which is related to the activation of pro-inflammatory and anti-inflammatory cytokines. Ulinastatin (Urinary trypsin inhibitor, UTI) is a serine protease inhibitor found in human urine or serum that inhibits the activation of human leukocyte elastase. This study examined the effect of UTI on the inflammation response in patients undergoing a gastrectomy. METHODS: Thirty patients scheduled to undergo a gastrectomy were divided into two groups as follows: Control group (untreated, n = 15) and UTI group (100,000 units of UTI were continuously injected intravenously for 2 hours, n = 15). Arterial blood was sampled before surgery (T0), 10 minutes after its onset (T1), at its end (T2), and 1 hour after surgery (T3) to measure the level of cytokines. RESULTS: Both the control and treatment groups had higher interleukin (IL)-6 levels at T2 and T3 than T0, and the level increased with time. However, the increase was smaller in the treatment group. The IL-8 levels were not activated significantly in any of the groups. CONCLUSIONS: UTI inhibits the secretion of IL-6, which is an inflammatory cytokine produced after a gastrectomy. This shows that UTI can decrease the inflammation reaction caused by surgical stress.

The Effects of Urinary Trypsin Inhibitor on the Outcomes of Severe Sepsis and Septic Shock Patients

PURPOSE: We purposed to determine the effects of urinary typsin inhibitor (ulinastatin) on the outcomes of severe sepsis and septic shock patients. METHODS: This is a prospective case control study of severe sepsis and septic shock patients who visited emergency department of university hospital from January 2005 to June 2008. For study group, 100,000 U of ulinastatin was initially infused and then additional infusions of ulinastatin were determined by the mean arterial pressure. We compared the predicted mortality and the actual in-hospital mortality between the ulinastatin group and the control group. We also compared the improvement of the SOFA score according to time between the groups. RESULTS: There were 43 patients in the ulinastatin group and 126 patients in the control group. The predicted mortality and the actual mortality of the ulinastatin group were 31.2% and 18.6%, respectively. The predicted and actual mortalities of the control group were 33.1% and 27.0%, respectively. The improvement of the SOFA score for the ulinastatin group was 6.8+/-3.9 and 5.0+/-4.5 at 0 and 24 hours (p<0.001), 6.5+/-3.7 and 3.9+/-4.3 at 0 and 48 hours (p<0.001) and, 6.3+/-3.6 and 3.0+/-4.1 at 0 and 72 hours (p<0.001). For the control group, the change of the SOFA score was 4.9+/-2.9 and 5.8+/-4.1 at 0 and 24 hours (p=0.003), 5.0+/-2.8 and 5.1+/-4.2 at 0 and 48 hours (p=0.760) and, 4.8+/-2.7 and 4.34.1 at 0 and 72 hours (p=0.105). CONCLUSION: The ulinastatin group showed significantly lower mortality than the predicted mortality and the ulinastatin group's SOFA score was improved in the early hospital days.

Purification and Characterization of a Protease Inhibitor from Fagopyrum tartaricum Gaertn Seeds and Its Effectiveness Against Insects / 中国生物化学与分子生物学报

Protease inhibitors, which are widely distributed in all types of life forms, are generally considered to be one of the most abundant proteins and a defense mechanism. A protease inhibitor from tartary buckwheat seeds (TBTI-Ⅱ ), with specific trypsin-inhibitory activity, was obtained by Resource Q anion-exchange chromatography and Superdex G 75 gel filtration. SDS-PAGE analysis indicated that the approximate molecular weight was 9.0 kD. Amino-acid analysis showed that the TBTI- Ⅱ was composed of 80 amino-acid residues with a high content of glutamate, aspartate and arginine. The inhibitor had high thermostability and retained 67.6% of its activity after heating at 100℃ for 10 min. The inhibition constant Ki was determined to be 1.01× 10-4 mol/L. It was demonstrated that the inhibitor was able to have an effect on the growth of cotton bollworm larva, after being fed with the artificial diets mixed with the target inhibitor. The present study indicates that the trypsin inhibitor from tartary buckwheat seeds could be a new potential anti-insect factor.

Effect of Prophylactic Administration of Trypsin Inhibitors on Pancreatic Islet Isolation of Adult Porcine / 中国普外基础与临床杂志

Objective To develop a method of adult porcine pancreatic islet isolation.Methods The tails of adult porcine pancreas were perfused through the pancreatic duct with 0.1% cold collagenase(type Ⅺ) and incubated at 38.5 ℃.The digested tissue was dispersed in 4 ℃ Hanks balanced salt solution(HBSS).The tissue suspensions were filtered through a 600 ?m mesh.The residual tissue was resuspended in cold HBSS,and put in the Ricordi’s chamber and shaken for 5 minutes,then filtered again.The isolated islets were divided into three groups: control group( n =14),Pefabloc(trypsin inhibitor, n =8) group and FOY(trypsin inhibitor, n =5) group.The collagenase solution of the Pefabloc and FOY group was supplemented with 1.0 mmol/L Pefabloc and FOY respectively. Results The islet yields of the Pefabloc group and FOY group 〔(11 848?3 530) islet/g pancreas and (14 496?3 693) islet/g pancreas〕 were significantly higher than that of the control group 〔(8 505?3 349) islet/g pancreas〕, P

Inhibitory effect of urinary trypsin inhibitor on IL-6 expression in severe acute pancreatitis in rats / 第三军医大学学报

Objective To explore the role of human urinary trypsin inhibitor(UTI) in severe acute pancreatitis (SAP). Methods UTI was highly purified from the urine of patients with acute pancreatitis by column chromatography. SAP models were established by injection of 5% sodium taurocholate(NaTc) at dose of 0.1 ml/100 g weight under pancreatic capsule. The interleukin 6(IL 6) level in each group was measured by ELISA. Results ① The highly purified UTI with high yield was harvested. ② SAP models were established successfully. ③ The level of IL 6 in SAP group was significantly higher than that in NC, but in UTI group, the increase was not obvious. Conclusion The alleviating effect of UTI on the pathological damage degree in rats with SAP is related to the inhibitive effect of UTI on the expression of IL 6.

The Study of Effect and Mechanism of Trypsin Inhibitor on Reperfusion Injury After Cerebral Focal Ischemia / 中国医师杂志

Objective To study the protective effect and molecular mechanism of trypsin inhibitor on reperfusion injury after cerebral ischemia. Methods The model of ischemia for 1h and repufusion for 24h of rat cerebrum was set by ligating MCAO described by zea-longa. 24 male rats were divided randomly into the sham operation group, the control group and trypsin inhibitor group. The presence of neurological function deficit was measured by Zea-Longa method, and the immunohistochemical staining and TUNEL were used to detect p53 protein expression and cell apoptosis in the brain tissues respectively. Results The score of neurological function deficit was zero in the sham operation group, 2 8?1 0 in the control group and 1 3?0 7 in trypsin inhibitor group. There were 12 3?2 5 p53 immunostaining positive cells in the control group and 5 5?1 3 in trypsin inhibitor group. The number of apoptotic cells was zero in the sham operation group, 7 6?1 0 in the control group, and 3 5?0 9 in trypsin inhibitor group. There was a significant difference in all above observing indices between the control group and trypsin inhibitor group(P

The effect of ulinastatin on blood fibrinolytic system and platelet function during cardiopulmonary bypass / 中华麻醉学杂志

Objective The aim of this study was to determine the effects of ulinastatin, a broad spectrum proteinase inhibitor, on fibrinolytic system and platelet function during open heart surgery performed with cardiopulmonary bypass (CPB) .Methods Twenty ASA Ⅰ-Ⅱpatients of both sexes undergoing cardiac surgery under CPB were randomly divided into two groups of ten patients: control group(C) and ulinastatin group (U). In group U patients received ulinastatin 12 000 U?2kg-1 . Half of the dose was given iv 10 min before CPB and the other half was added to the priming solution. In group C patients received normal saline instead of ulinastatin. Blood samples were taken before CPB (T1 ) , 30 min after CPB was started (T2), at the end of CPB (T3), 2 h and 4 h after CPB(T4 , T5) for determination of plasma levels of D-Dimer, ?-granule membrane protein-140 (GMP-140), thromboxane B2 (TXB2) and 6-Keto-prostaglandin F1? (6-Keto-PGF1?) .Results The demographic data, aortic cross-clamping time, CPB time and duration of operation were comparable between the two groups. The plasma levels of D-Dimer, GMP-140, TXB2 and TXB2/6-Keto-PGFl? were significantly increased at T2 , T3 and T4 as compared with the baseline (T1 ) in both groups, but the increase was significantly larger in group C than in group U(P