Strain difference of susceptibility to caerulein-induced acute pancreatitis in mice

AIM: To study the difference of susceptibility to caerulein-induced acute pancreatitis (AP) among the mice of C57BL/6J, BALB/c and ICR strains.METHODS: Two-month-old female mice of C57BL/6J, BALB/c and ICR strains (12 mice for each strain) were divided into control group (n=6) and experimental group (n=6), respectively. The mice were intraperitoneally injected with caerulein (50 μg/ kg) in 1 h interval for 7 serial injections in total. The mice in control group were treated with saline according to the same procedure in experimental group. The blood samples were collected at 0 h, 3 h, 6 h, 9 h, 12 h and 24 h after the first injection of caerulein or saline for plasma α-amylase and lipase assays. The mice were sacrificed 24 h after AP induction, and the pancreatic tissues were harvested for further investigating the pathological changes and expression of inflammatory factors.RESULTS: After AP induction, the mice of BALB/c and ICR strains demonstrated more dramatic increase in plasma α-amylase activity and lipase activity than those of C57BL/6J mice. C57BL/6J mice showed milder morphological changes and lower expression of inflammatory factors in pancreata than those of BALB/c and ICR mice.CONCLUSION: The mice of C57BL/6J strain have less susceptibility to caerulein-induced AP than that of BALB/c and ICR mice.     

Influence of exogenous somatostatin on pancreatic blood flow in normal rats or rats with acute necrotizing pancreatitis

AIM:To investigate the influence of exogenous somatostatin (stilamin) on pancreatic blood flow in normal rats or rats with acute necrotizing pancreatitis (ANP).METHODS:Pancreatic blood flow (PBF) was detected with computerized tissue blood flowmeter and rats with ANP were triggered with sodium taurocholate. Metabolites of eicosanoids in plasma were determined with radioimmunoassay. Other laboratory tests including histopathologic observation under optical or electron microscope were used. RESULTS:There was a significant decrease in PBF in normal rats after stilamin administration in comparison with that before use of the drug. There was significant decrease in PBF after onset of ANP, but, compared with that in ANP group, significant increase was shown in SS(stilamin)+ANP group. Plasma thromboxin-B₂(TXB₂) in ANP group at 6 hours after ANP was significantly higher, with increase of 4.5 times, than that in Sham(sham operated) group while TXB₂, detected each time during the course of ANP, significantly decreased in SS+ANP group. 6-Keto-prostagland in F_1α(6-Keto-PGF_1α) at 6 h after ANP was significantly higher, and the ratio of TXB ₂／6-Keto-PGF_1α, significantly lower in SS+ANP group than that in ANP group. Lessened necrosis of acinar cells, along with much fewer microthrombi in microvessels in SS+ANP group, was shown by pathologic scoring or electron microscope than that in ANP group.CONCLUSION:Administration of exogenous somatostatin leads to the decrease in PBF in physiological setting but it attenuates pancreatic ischemia in SS+ANP group, which may be attributed to correction of abnormal metabolism of eicosanoids, improvement of pancreatic microcirculation and cytoprotection of acinar cells as well.     

Effect of grape seed proanthocyanidin on cisplatin-induced nephrotoxicity in mice

AIM:To study the protective effect of grape seed proanthocyanidin (GSP) on nephrotoxicity induced by cisplatin (CP) in mice. METHODS:Male C57 mice were randomly divided into 4 groups: control group (N group, n=10), CP group (intraperitoneal injection of CP at dose of 20 mg/kg, n=20), GSP group (intragastric administration of GSP at dose of 500 mg/kg, n=15) and CP+GSP group (intragastric administration of GSP 30 min prior to intraperitoneal injection of CP and intragastric administration of the same dose of GSP 72 h later, n=20). On the 5th day after CP treatment, blood and kidney samples were collected. The renal pathological changes were examined by HE staining. The protein expression of glucose-regulated protein 78 (GRP78) and phosphorylated extracellular signal-regulated kinase (p-ERK) was examined by Western blotting and immunohistochemical staining. RESULTS:Renal index, blood urea nitrogen and serum creatinine were significantly higher in CP group than those in N group. The renal tissues were heavily damaged in CP group. The protein expression of GRP78 and p-ERK increased in CP group compared with N group. GSP treatment alleviated the increase in the renal index, blood urea nitrogen and serum creatinine. The damages of the renal tissues were attenuated. The protein expression of GRP78 and p-ERK was obviously reduced in CP+GSP group compared with CP group. CONCLUSION:GSP prevents kidney from CP-induced damage by suppression of endoplasmic reticulum stress.     

Reduction of inflammatory-related factor expression in experimental acute pancreatitis in Egr-1 knockout mice

AIM: To observe the effects of Egr-1 gene knockout on the expression of inflammatory-related factors in pancreatic tissue in a mouse acute pancreatitis model. METHODS: The experimental pancreatitis was induced by high-dose of cearulein in wildtype mice and Egr-1 knockout mice. The pancreatitis indexes, such as serum amylase, pancreata edema, and myeloperoxidase(MPO) levels in pancreata and lungs were recorded. The mRNA levels of tissue factor(TF), plasminogen activator inhibitor(PAI-1), monocyte chemoattractant protein(MCP-1), Gro-1, IL-6 and ICAM-1 were measured by quantitative PCR. RESULTS: Contrary to wildtype mice, typical pancreatitis was not induced by high-dose cearulein in the Egr-1 knockout mice, not only markedly reduced edema in pancreata and lungs, but decreased MPO levels in lungs as well were found. Furthermore, the mRNA of TF, PAI, MCAP, ICAM-1 and IL-6 in pancreata were significantly decreased in Egr-1 knockout mice. CONCLUSION: The severity of pancreatitis and lung damage is ameliorated in Egr-1 knockout mice stimulated by high-dosage of cearulein, which was probably mediated by decreasing expression of inflammatory-related factors in pancreata, such as TF, PAI, MCP-1, ICAM-1 and IL-6.     

葡萄子油及原花青素研究与开发利用

综述了葡萄子油、葡萄子原花青素等天然产物的提取与保健作用的研究进展,原花青素的生物活性及药理研究的现状,葡萄子油成分分析方法和葡萄子,提取物中原花青素含量测定的方法;介绍了用超临界CO2萃取法制取高质量的葡萄子油和应用夹带剂提取葡萄子中的原花青素的方法。分析了葡萄子的综合开发的应用前景。     

A one-stage model of experimental acute necrotizing pancreatitis in rats

AIM: To establish a one-stage model of experimental acute necrotizing pancreatitis (ANP) in rats characterized by the simplicity of performance and a high degree of repeatability. METHODS: ANP modeling in rats was performed based on modification of the ligation model as follows: synthetic material ligature using an atraumatic needle was performed to capture pancreatic gland ducts and marginal duodenum vessels. Ligature tips were exteriorized to the abdominal wall, and the ligature was skinned over to avoid catching intestine loops. Pancreatic macroscopic appearance and histological changes were observed. Blood biochemical and hemostatic indicators were also determined. RESULTS: Laboratory analysis of rats with experimental ANP showed a pattern of disturbances similar to that observed during pancreatic necrosis in humans as soon as the first day. General blood analysis revealed enhanced leukocytosis and alterations in leukogram characteristics, indicating acute inflammation. Serum levels of amylase, aspartate aminotransferase and creatinine significantly increased (P<0.05). Hemostatic indicators showed alterations indicating formation of disseminated intravascular coagulation, and signs of endotoxicosis were observed. These typical pancreatic necrosis patterns of disturbances were validated by the results of histological investigation. CONCLUSION: Histological changes and laboratory indicators confirm the development of a suitable model of ANP.     

葡萄组培苗耐盐性研究

对中国野生葡萄4个种的6个株系、2个种间杂种、5个鲜食葡萄品种、1个酿酒葡萄品种和5个砧木品种通过组织培养获得试管苗,以不同浓度的NaCl作为盐胁迫因子,分别加入生根培养基中进行耐盐筛选。结果表明,在盐胁迫的50d内,试管苗的侧芽萌发倍数、增殖系数、生根率、生根数目均随着盐浓度的升高而降低,试管苗的受害指数、胁迫敏感指数随着盐浓度的升高而增大。在0.6%NaCl胁迫下,随着盐胁迫时间的延长,受害率和受害指数呈上升趋势。将供试的19个材料耐盐性分为4个类型:耐盐葡萄类型,包括酿酒葡萄EndFine、燕山葡萄燕山-1、1-1-6(88-110)、莫里莎无核和克瑞森无核;较耐盐葡萄类型,包括红地球、1-1-8(88-110)、户太8号、山葡萄通化-3和秋葡萄平利-7;盐较敏感葡萄类型,包括京秀、101-14MG、3309C和蘡奠葡萄泰山-1、安林-3;盐敏感葡萄类型,包括SO4、1103P、110R和山葡萄华县-47。     

葡萄梗疣形成过程及内部结构观察

用扫描电子显微镜和光学显微镜观察了5个葡萄品种小果柄上疣的大小、形成过程、内部形态、组织结构与病菌的侵染情况。发现抗性较强的北醇,小果柄上的疣小、形成较早,发育速度较慢,破裂后内部结构层次清晰,基部木栓形成层明显,抗性差的红地球则相反。推测该组织可能与葡萄果实病害病原菌的侵染有关。     

不同葡萄砧木对矢富罗莎葡萄嫁接苗光合作用的影响

在避雨条件下,以盆栽试验观察不同砧木(巨峰、1202、5BB、110R、99R、华佳8号、8B和SO4)对矢富罗莎葡萄叶片光合日变化的影响。结果表明,不同葡萄砧木矢富罗莎嫁接苗叶片净光合速率(Pn)的日变化基本呈双峰型曲线。叶片净光合速率日变化存在差异,99R、华佳8号和1202砧穗组合嫁接苗叶片的净光合速率(Pn)明显高于其他组合,最高峰值(11:00时)分别高出对照128.8%、106.0%和77.3%;叶片气孔导度(Gs)值1202、110R、99R和华佳8号组合较高,9:00时分别达到对照的8.5、8.0、7.5和6.5倍;99R、华佳8号、110R和1202组合的叶片蒸腾速率(Tr)显著高于其他组合和对照,巨峰和8B组合居中。     

不同化学物质打破葡萄、桃休眠的研究

以保护地栽培的葡萄、桃为试材,对氰氨类、细胞分裂素、赤霉酸和KNO3打破休眠的效果进行了研究。结果表明,1.5%单氰氨、11%石灰氮可明显促使葡萄提前萌芽,提早着色9-10d;50mg/LGA3和100mg/L6-BA对打破休眠基本无效。TDZ与单氰氨混用提高萌芽率的效果最好。在2个葡萄品种上,均没有观察到单氰氨或石灰氮处理对果穗或果实品质性状的不良影响。在华光油桃上,没有观察到化学物质对提早开花或成熟有明显的作用。对葡萄和桃上应用化学物质打破休眠的有关问题进行了讨论。     

Influence of SDD and caecostomy/colonic irrigation on gut endotoxin/bacteria translocation following acute severe pancreatitis

AIM: To observe the influence of the selective decontamination of the digestive tract (SDD) and caecosomy/colonic irrigation on gut endotoxin/bacteria translocation following acute severe pancreatitis (ASP). METHODS: Twenty three pigs weighing 16-22 kg were divided into four groups. Group I (n=5): sham-control; Group Ⅱ (n=6): ASP-control; Group Ⅲ (n=6): gntamicin [(8.55×10⁵±5.70×10⁴)units/time] and nystatin [(1.37×10⁵±9.00×10³)units/time]were fed orally every 8 h for 1 week before the induction of ASP; Group Ⅳ (n=6): caecostomy was performed before the induction of ASP. ASP was induced by infecting 1 mL/kg BW of combined solution of 5% sodium taurocholate and (8-10)×10⁶ BAEE units/L of trypsin into pancreas via pancreatic duct. Systemic plasma endotoxin levels were quantified by the chromogenic limulus amebocyte lysate (LAL) technique. Specimens of tissue from mesenteriolum and mesocolon lymph nodes, lung, lymph nodes in hilus pulmonis, pancreas and the samples of both portal and systemic blood were collected before and at 72 h following ASP and cultured for aerobic as well as anaerobic bacteria growth. Positive specimens were subcultured and the bacteria identified by standard procedure. RESULTS: Preventive SDD not only effectively reduced the amount of bacteria in stool (P<0.01), but also significantly reduced the levels of plasma endotoxin and the magnitude of bacteria translocation to the portal and systemic blood and the remote organs and tissues, for instance, mesenteriolum and mesocolon lymph nodes, lung, lymph nodes in hilus pulmonis, pancreas. Early caecostomy/colonic irrigation also significantly reduced the levels of translocated origin-endotoxin and bacteria after ASP. CONCLUSIONS: SDD and caecostomy/colonic irrigation effectively reduce the levels of plasma endotoxin and the magnitude of bacteria translocation to the portal and systemic blood and the remote organ, especially the latter will be of a great importance in the future clinical practice.     

葡萄愈伤组织制备原生质体的影响因素研究

【目的】为了获得高质量的葡萄原生质体,【方法】以"鄞红"葡萄茎段愈伤组织为材料,对葡萄原生质体制备过程中的若干影响因子,如酶解液成分、甘露醇浓度、酶解时间、离心条件等方面进行了研究。【结果】0.5 mol.L-1的甘露醇+1%纤维素酶+0.025%～0.05%果胶酶为最佳的酶解液组成;酶解14 h、120×g离心3 min条件下原生质体数量和质量最佳。【结论】研究获得的高质量葡萄原生质体,为下一步的植株再生及葡萄品种遗传改良提供了良好的材料。     

Role of stress in the lung injury caused by acute hemorrhagic necrotizing pancreatitis in rats

AIM:To study the relation ship between stress and lung injury caused by acute hemorrhagic necrotizing pancreatitis(AHNP) through AHNP model.METHODS:The AHNP model was made by using 5% sodium taurocholate retrograded injection into biliopancreatic duct in SD rats. Those rats were divided into three groups randomly, from A to C, the A group undertook sham operations, the B group was made into AHNP model, and the C group was given Metyrapone. The level of corticosteroid, CRP and amylase in serum had been observed. The lung and pancreas histological examinations were also performed.RESULTS: In C group, the level of corticosteroid, CRP and amylase were much lower than those in B group. The grade of lung and pancreas injury were also lower than those in B group(P＜0.05).CONCLUSION:Stress lays an important role in the lung injury caused by AHNP. Using anti-stress drugs can inhibit this course and meliorate the lung injury.     

Influence of fluid resuscitation on pancreatic microthrombosis in severe acute pancreatitis

AIM: To make evaluation on fluid resuscitation with either hypertonic saline (HS) or dextran 40 (Dx) on pancreatic microthrombi and dysfunction of microcirculation in rats with severe acute pancreatitis (SAP).METHODS: SD rats were allocated into 4 groups randomly,ie.SAP group,HS group,Dx group,which respectively received normal saline (NS,4 mL/kg),HS (4 mL/kg),Dx (4 mL/kg) for 2 h by the tail intravenous injection consecutively after being made as SAP animal models,and operate sham group (OS).12 h after the operation,all animals were blooded to assess the serum amylase levels,plasma D-dimer,von Willebrand factor and GMP-140 levels.The amount of ascites was measured and the samples of the pancreas were collected for pathologic examination under light microscopy as well as transmission electron microscope.The numbers of pancreatic microthrombi were also counted with microscopy.RESULTS: (1) 12 h later when the rats were sacrificed,the survival rate in SAP group was the lowest,significantly lower than that in the 2 fluid resuscitation groups (P<0.05).However,no significant difference was found between the latter 2 groups though the survival numbers.(2) The numbers of microthrombi in the pancreas and the histological scoring of pancreatic injury were significantly fewer in the 2 fluid resuscitation groups than those in SAP group (P<0.05) though no significant difference was revealed between the 2 treated groups.The amounts of ascites,serum amylase,plasma D-dimer,von Willebrand factor and GMP-140 showed significant improvement in HS or Dx group compared to those in SAP group(P<0.05,P<0.01).However,no significant difference was found between the 2 fluid resuscitation groups.CONCLUSION: Fluid resuscitation with HS or Dx,if applied at the early stage,increases survival rate with lessened histological injury,fewer microthrombi and improved coagulation parameters including D-dimer,von Willebrand factor and GMP-140.Either hypertonic saline (HS) or dextran 40 (Dx) demonstrates the same improvement in the relevant laboratory parameters in SAP.     

两种抗生素对葡萄试管苗芽孢杆菌抑制效应的研究

用医用头孢唑啉钠、青霉素钠两种抗生素,以不同的处理方法和不同的浓度处理在继代培养中被芽孢杆菌污染的葡萄试管苗。结果表明：对芽孢杆菌的抑制作用,青霉素钠优于头孢唑啉钠,添加处理优于浸泡处理,并且添加处理中培养基灭菌前加与灭菌后加抑制效果差异极显著。培养基中的药剂浓度在0～50mg／L之间能显著抑制芽孢杆菌的发生,并能促进植株生长;浓度大于50mg／L,对植株的生长有明显的抑制作用。所以灭菌前添加青霉素钠溶液,配成药剂浓度为50me／L的改良B5培养基有利于防止芽孢杆菌的发生,并能促进植株的生长。     

Effect of experimental acute necrotizing pancreatitis on sodium and L-type calcium current in rat cardiomyocytes

AIM: To study the effect of experimental acute necrotizing pancreatitis (ANP) on sodium and L-type calcium current in rat cardiomyocytes. METHODS: I_Na and I_Ca-L were recorded using whole cell patch-clamp techniques from left ventricular myocytes in ANP model established by retrograde injection of 3.5% sodium taurocholate 2.5 mL/kg into pancreatic duct. RESULTS: Peak I_Na current density (at -30 mV) was significantly reduced in ANP [(12.45±2.26)pA/pF,n=16] compared with sham [(25.32±3.31)pA/pF,n=14], P<0.01; I_Ca-L current density (at +10 mV) was also significantly reduced in ANP [(3.63±0.65)pA/pF,n=16] compared with sham [(5.46±1.03)pA/pF,n=12], P<0.05. CONCLUSIONS: There were changes in both I_Na and I_Ca-L in cardiomyocytes of ANP. These changes may underlie the altered excitability and abnormally short transmembrane action potentials and repolarization of cardiomyocytes, thus contributing to arrhymias in ANP.     

Synergistic and attenuating effects of Qiongyugao on pancreatic cancer mice with chemotherapy

AIM: To observe the synergistic and attenuating effects of Qiongyugao on pancreatic cancer mice with chemotherapy and to explore its possible mechanism. METHODS: C57BL/6 mice (n=50) were randomly divided into normal group, model group, gemcitabine treatment group, Qiongyugao treatment group, and combined treatment (gemcitabine+Qiongyugao) group. The pancreatic cancer mouse xenograft model was established by subcutaneous inoculation with pancreatic cancer Panc02 cells among the mice in model group and 3 treatment groups. Normal saline, gemcitabine, Qiongyugao and gemcitabine plus Qiongyugao were also used accordingly. The tumor growth curve, blood routine and tumor markers were observed in each group. Furthermore, flow cytometry was used to detect the expression of Th1/Th2 and Th/Treg in peripheral blood of the mice. RESULTS: Subcutaneous vaccination with Panc02 cells successfully established a transplanted tumor model of pancreatic cancer. Compared with model group, Qiongyugao, gemcitabine and the combination of the 2 drugs inhibited tumor growth (P<0.05), and decreased the concentration of carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA) (P<0.05). Qiongyugao decreased the ratio of Th1/Th2 and Th17/Treg, and increased the white blood cell (WBC), red blood cell (RBC) and platelet (PLT) counts and hemoglobin (HGB) content in gemcitabine treated mice (P<0.01). CONCLUSION: Qiongyugao may have a certain therapeutic effect on pancreatic cancer and produce certain synergistic and attenuating effects on gemcitabine chemotherapy.     

葡萄果皮花色素的提取及其理化性质

以红地球葡萄品种为试材,研究了葡萄果皮中花色素的提取方法及色素理化性质。结果表明,1%盐酸-无水甲醇的提取葡萄果皮色素效率最高,其次是95%乙醇-4.7%盐酸(v/v=85/15)、1%盐酸-无水乙醇;以1%盐酸-无水乙醇为提取液,葡萄果皮质量与提取液的料液比宜为1∶5～1∶10(m∶v,g/mL)。光谱特性分析表明,葡萄皮花色素属于花色素苷类,其稳定性受溶液的pH值影响最大,当溶液pH≥9时,特征光谱消失。葡萄花色素具有一定的热稳定性,但在光下降解速度加快,受热后在光下降解更快;Fe3+对葡萄花色素的不良影响大于Zn2+、Ca2+;高浓度的蔗糖、果糖对花色素有一定的护色效应,而葡萄糖对其影响不明显;维生素C、苯甲酸钠对花色素有不良影响。