坏死性胰腺炎手术前后处理的几点体会

坏死型胰腺炎,来势凶猛,病情复杂,需从多方面进行救治。我院近五年共收治9例,7例治愈。现结合本组病例,就休克的防治,术后胰床引流及完全胃肠外营养三个问题,浅谈我们的体会。一、休克的防治急性胰腺炎的基本病理特征为大量胰酶释放导致自体消化坏死。因胰激酶等血管活体物质的大量释放,使末稍血管扩张,血管通透性增加,有效血容量减少。

Tyrosine kinase of insulin-like growth factor receptor as target for novel treatment and prevention strategies of colorectal cancer

AIM： To investigate the antineoplastic potency of the novel insulin-like growth factor 1 receptor （IGF-1R） tyrosine kinase inhibitor （TKI） NVP-AEW541 in cell lines and primary cell cultures of human colorectal cancer （CRC）. METHODS： Cells of primary colorectal carcinomas were from 8 patients. Immunostaining and crystal violet staining were used for analysis of growth factor receptor protein expression and detection of cell number changes, respectively. Cytotoxicity was determined by measuring the release of the cytoplasmic enzyme lactate dehydrogenase （LDH）. The proportion of apoptotic cells was determined by quantifying the percentage of sub-G1 （hypodiploid） cells. Cell cycle status reflected by the DNA content of the nuclei was detected by flow cytometry. RESULTS： NVP-AEW541 dose-dependently inhibited the proliferation of colorectal carcinoma cell lines and primary cell cultures by inducing apoptosis and cell cycle arrest. Apoptosis was characterized by caspase-3 activation and nuclear degradation. Cell cycle was arrested at the G1/S checkpoint. The NVP-AEW541-mediated cell cycle-related signaling involved the inactivation of Akt and extracellular signal-regulated kinase （ERK） 1/2, the upregulation of the cyclin-dependent kinase inhibitors p21^waf1/CIP1 and p27^kjp1, and the downregulation of the cell cycle promoter cyclin D1. Moreover, BAX was upregulated during NVP-AEW541-induced apoptosis, whereas Bcl-2 was downregulated. Measurement of LDH release showed that the antineoplastic effect of NVP-AEW541 was not due to general cytotoxicity of the compound. However, augmented antineoplastic effects were observed in combination treatments of NVP-AEW541 with either 5-FU, or the EGFR-antibody cetuximab, or the HMG-CoA-reductase inhibitor fluvastatin. CONCLUSION： IGF-1R-TK inhibition is a promising novel approach for either monoor combination treatment strategies of colorectal carcinoma and even for CRC chemoprevention.

The roles of MAPKs in disease

MAP kinases transduce signals that are involved in a multitude of cellular pathways and functions in response to a variety of ligands and cell stimuli. Aberrant or inappropriate functions of MAPKs have now been identified in diseases ranging from cancer to inflammatory disease to obesity and diabetes. In many cell types, the MAPKs ERK1/2 are linked to cell proliferation. ERK1/2 are thought to play a role in some cancers, because mutations in Ras and B-Raf, which can activate the ERK1/2 cascade, are found in many human tumors. Abnormal ERK1/2 signaling has also been found in polycystic kidney disease, and serious developmental disorders such as cardio-facio-cutaneous syndrome arise from mutations in components of the ERK1/2 cascade. ERK1/2 are essential in well-differentiated cells and have been linked to long-term potentiation in neurons and in maintenance of epithelial polarity. Additionally, ERK1/2 are important for insulin gene transcription in pancreatic beta cells, which produce insulin in response to increases in circulating glucose to permit efficient glucose utilization and storage in the organism. Nutrients and hormones that induce or repress insulin secretion activate and/or inhibit ERK1/2 in a manner that reflects the secretory demand on beta cells. Disturbances in this and other regulatory pathways may result in the contribution of ERK1/2 to the etiology of certain human disorders.

Presence of CCK-A, B receptors and effect of gastrin and cholecystokinin on growth of pancreatobiliary cancer cell lines

AIM: To investigate the effects of gastrin and cholecystokinin (CCK) and their specific antagonists on the growth of pancreatic and biliary tract cancer cell lines. METHODS: Five pancreatic and 6 biliary cancer cell lines with 2 conrtol cells were used in this study. Cell proliferation study was done using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) test and direct cell count method. Reverse transcription-polymerase chain reaction (RT-PCR) and slot blot hybridization were performed to examine and quantify the expression of hormonal receptors in these cell lines. RESULTS: SNU-308 showed a growth stimulating effect by gastrin-17, as did SNU-478 by both gastrin-17 and CCK-8. The trophic effect of these two hormones was completely blocked by specific antagonists (L-365, 260 for gastrin and L-364, 718 for CCK). Other cell lines did not respond to gastrin or CCK. In RT-PCR, the presence of CCK-A receptor and CCK-B/gastrin receptor mRNA was detected in all biliary and pancreatic cancer cell lines. In slot blot hybridization, compared to the cell lines which did not respond to hormones, those that responded to hormones showed high expression of receptor mRNA. CONCLUSION: Gastrin and CCK exert a trophic action on some of the biliary tract cancers.

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

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

Pancreatic function testing: Here to stay for the 21st century

The diagnosis of Chronic Pancreatitis （CP） is based on the detection of abnormal structure or function of the diseased pancreas. The pancreatic function tests more accurately determine the presence of CP than tests of structure, especially for early stage disease. The function tests can be divided into two categories： non- invasive and invasive. The invasive ＂tube＂ tests can reliably detect mild, early CP, but are only available at a few referral centers and tend to be poorly tolerated by patients. The non-invasive tests are easy to obtain, but tend to perform poorly in patients with early, mild disease. Therefore, no one test is useful in all clinical situations, and a detailed understanding of the rational, pathophysiologic basis, strengths, and limitations of various tests is needed. This review highlights the role of various pancreatic function tests in the diagnosis of CP including fecal fat analysis, fecal elastase, fecal chymotrypsin, serum trypsin, the secretin stimulation test, the cholecystokinin （CCK） stimulation test, the combined secretin-CCK stimulation test, the intraductal and endoscopic secretin stimulation tests, and the functional magnetic resonance imaging of the pancreas after secretin stimulation.

牛磺熊去氧胆酸对DSS诱导大鼠溃疡性结肠炎的治疗作用及可能机制

目的:探究牛磺熊去氧胆酸(TUDCA)对DSS诱导的大鼠溃疡性结肠炎的治疗作用及可能机制。方法:选取健康成年雄性Sprague-Dawley(SD)大鼠30只,随机分为正常组、模型组、治疗组,每组10只。模型组和治疗组给予葡聚糖硫酸钠(dextran sulphate sodium,DSS)诱导大鼠溃疡性结肠炎模型。正常组和模型组给予生理盐水灌胃,治疗组给予TUDCA灌胃,治疗7 d后处死大鼠,观察大鼠的一般情况及结肠组织病理改变。比较三组DAI、HI评分。Western blot检测大鼠结肠组织GRP78、PERK、eIF2α、ATF4、CHOP蛋白表达水平。结果:治疗组大鼠一般情况及结肠组织病理情况均优于模型组。模型组、治疗组DAI评分均明显高于正常组(P<0.05),而治疗组低于模型组,差异有统计学意义(P<0.05)。模型组、治疗组HI评分均明显高于正常组(P<0.05),而治疗组低于模型组,差异有统计学意义(P<0.05)。模型组、治疗组结肠组织GRP78、PERK、eIF2α、ATF4、CHOP蛋白表达水平均明显高于正常组(P<0.05),而治疗组低于模型组,差异有统计学意义(P<0.05)。结论:TUDCA对DSS诱导的大鼠溃疡性结肠炎具有治疗作用,其机制可能是通过抑制PERK-eIF2α-ATF4-CHOP通路发挥分子生物学作用。

Insights into erlotinib action in pancreatic cancer cells using a combined experimental and mathematical approach

AIM:To gain insights into the molecular action of erlotinib in pancreatic cancer (PC) cells. METHODS:Two PC cell lines, BxPC-3 and Capan-1, were treated with various concentrations of erlotinib, the specific mitogen-activated protein kinase kinase (MEK) inhibitor U0126, and protein kinase B (AKT) inhibitor XIV. DNA synthesis was measured by 5-bromo-2'-deoxyuridine (BrdU) assays. Expression and phosphorylation of the epidermal growth factor receptor (EGFR) and downstream signaling molecules were quantified by Western blot analysis. The data were processed to calibrate a mathematical model, based on ordinary differential equations, describing the EGFRmediated signal transduction. RESULTS:Erlotinib significantly inhibited BrdU incorporation in BxPC-3 cells at a concentration of 1 mol/L, whereas Capan-1 cells were much more resistant. In both cell lines, MEK inhibitor U0126 and erlotinib attenuated DNA synthesis in a cumulative manner, whereas the AKT pathway-specific inhibitor did not enhance the effects of erlotinib. While basal phosphorylation of EGFR and extracellular signal-regulated kinase (ERK) did not differ much between the two cell lines, BxPC-3 cells displayed a more than five-times higher basal phospho-AKT level than Capan-1 cells. Epidermal growth factor (EGF) at 10 ng/mL induced the phosphorylation of EGFR, AKT and ERK in both cell lines with similar kinetics. In BxPC-3 cells, higher levels of phospho-AKT and phospho-ERK (normalized to the total protein levels) were observed. Independent of the cell line, erlotinib efficiently inhibited phosphorylation of EGFR, AKT and ERK. The mathematical model successfully simulated the experimental findings and provided predictions regarding phosphoprotein levels that could be verified experimentally. CONCLUSION:Our data suggest basal AKT phosphorylation and the degree of EGF-induced activation of AKT and ERK as molecular determinants of erlotinib efficiency in PC cells.

Insulin-like growth factor binding protein-5 influences pancreatic cancer cell growth

AIM： To investigate the functional significance of insulin-like growth factor binding protein-5 （IGFBP-5） overexpression in pancreatic cancer （PaC）.METHODS： The effects of IGFBP-5 on cell growth were assessed by stable transfection of BxPC-3 and PANC-1 cell lines and measuring cell number and DNA synthesis. Alterations in the cell cycle were assessed by flow cytometry and immunoblot analyses. Changes in cell survival and signal transduction were evaluated after mitogen and phosphatidylinositol activated protein kinase 3-kinase （PI3K） inhibitor treatment.RESULTS： After serum deprivation, IGFBP-5 expression increased both cell number and DNA synthesis in BxPC-3 cells, but reduced cell number in PANC-1 cells. Consistent with this observation, cell cycle analysis of IGFBP-5-expressing cells revealed accelerated cell cycle progression in BxPC-3 and G2/M arrest of PANC-1 cells. Signal transduction analysis revealed that Akt activation was increased in BxPC-3, but reduced in PANC-1 cells that express IGFBP-5. Inhibition of PI3K with LY294002 suppressed extracellular signal-regulated kinase-1 and -2 （ERK1/2） activation in BxPC-3, but enhanced ERK1/2 activation in PANC-1 cells that express IGFBP-5. When MEK1/2 was blocked, Akt activation remained elevated in IGFBP-5 expressing PaC cells; however, inhibition of PI3K or MEK1/2 abrogated IGFBP-5-mediated cell survival.CONCLUSION： These results indicate that IGFBP-5 expression affects the cell cycle and survival signal pathways and thus it may be an important mediator of PaC cell growth.

Toxic Dinoflagellate Alexandrium tamarense Induces Oxidative Stress and Apoptosis in Hepatopancreas of Shrimp(Fenneropenaeus chinensis)

This study investigated the inductive effect ofAlexandrium tamarense, a toxic dinoflagellate producing paralytic shell- fish poison, on oxidative stress and apoptosis in hepatopancreas of Chinese shrimp, Fenneropenaeus chinensis. The individuals of E chinensis were exposed to 200 and 1000 cells mL-1 of A. tamarense with their superoxide dismutase （SOD）, glutathione S-transferase （GST） activities, malonyldialdehyde （MDA） concentration, and caspase gene （FcCasp） expression in hepatopancreas determined at 12, 24, 48, 72 and 96 h. In addition, apoptosis in hepatopancreas of E chinensis at 96 h after exposure was determined through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay. The hepatopancreatic SOD and GST activities of F. chinensis exposed to 1000 cells mL-1 ofA. tamarense showed a bell-shaped response to exposure time. The hepatopancreatic MDA concentration ofF. chinensis exposed to 1000 cellsmL-1 ofA. tamarense increased gradually from 48 to 96h, and such a trend corresponded to the decrease of GST activity. The hepatopancreatic FcCasp transcript abundance of F. chinensis exposed to 1000 cells mL-1 ofA. tamarense was positively and linearly correlated to MDA concentration. Results of TUNEL assay showed that exposure to 1000 cells mL-1 of A. tamarense induced apoptosis in the hepatopancreas of E chinensis. Our study revealed that A. tamarense exposure influenced the antioxidative status ofF. chinensis and caused lipid peroxidation and apoptosis in the hepatopancreas of shrimp.

On the Role of Intestinal Microbiota in Patients with Cognitive Decline

Objective： The association between gut microbiota composition and biomarkers of immune activation and inflammation was assessed in the elderly. Patients： Serum inflammation markers of fifty-five outpatients （29 females, 26 males, aged 78 ＋ 8.5 years） were analyzed. Stool specimens and thus data on gut microbiota were available from a subgroup of 23 individuals （9 females and 14 males）. Results： Global cerebral atrophy was found in all magnet resonance tomography scans. Mean mini-mental-score examination in Alzheimer＇s disease patients was 18.8 ± 7.1, in patients with mild cognitive impairment 27.8 ± 1.5. Serum neopterin concentrations correlated with concentrations of fecal S100A12 （p 〈 0.001） and cq-antitrypsin （p 〈 0.05）. Faecalibacterium prausnitzii correlated with MMSE （p 〈 0.05）, with Akkermansia muciniphila （p 〈 0.01） and with serum neopterin （p 〈 0.05）. Fecal zonulin correlated inversely with Clostridium cluster I （p 〈 0.02）. Conclusions： Our results underline earlier in vitro and animal studies that cognitive decline associates with age-related changes in the intestinal microbiota and neuroinflammation. However, only correlational evidence can be reported, and a causative relationship still has to be demonstrated.

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

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

Baicalin attenuates high fat diet-induced insulin resistance and ectopic fat storage in skeletal muscle,through modulating the protein kinase B/Glycogen synthase kinase 3 beta pathway

Insulin resistance is the pathophysiological basis of many diseases.Overcoming early insulin resistance highly significant in prevention diabetes,non-alcoholic fatty liver,and atherosclerosis.The present study aimed at evaluating the therapeutic effects of baicalin on insulin resistance and skeletal muscle ectopic fat storage in high fat diet-induced mice,and exploring the potential molecular mechanisms.Insulin resistance in mice was induced with a high fat diet for 16 weeks.Animals were then treated with three different doses of baicalin(100,200,and 400 mg·kg^(-1)·d^(-1)for 14 weeks.Fasting blood glucose,fasting serum insulin,glucose tolerance test(GTT),insulin tolerance test(ITT),and skeletal muscle lipid deposition were measured.Additionally,the AMP-activated protein kinase/acetyl-CoA carboxylase and protein kinase B/Glycogen synthase kinase 3 beta pathways in skeletal muscle were further evaluated.Baicalin significantly reduced the levels of fasting blood glucose and fasting serum insulin and attenuated high fat diet induced glucose tolerance and insulin tolerance.Moreover,insulin resistance was significantly reversed.Pathological analysis revealed baicalin dose-dependently decreased the degree of the ectopic fat storage in skeletal muscle.The properties of baicalin were mediated,at least in part,by inhibition of the AMPK/ACC pathway,a key regulator of de novo lipogenesis and activation of the Akt/GSK-3β pathway,a key regulator of Glycogen synthesis.These data suggest that baicalin,at dose up to 400 mg·kg^(-1)·d^(-1),is safe and able to attenuate insulin resistance and skeletal muscle ectopic fat storage,through modulating the skeletal muscle AMPK/ACC pathway and Akt/GSK-3β pathway.

Molecular variation and evolution of the tyrosine kinase domains of insulin receptor IRa and IRb genes in Cyprinidae

The insulin receptor （IR） gene plays an important role in regulating cell growth, differentiation and development. In the present study, DNA sequences of insulin receptor genes, IRa and IRb, were amplified and sequenced from 37 representative species of the Cyprinidae and from five outgroup species from non-cyprinid Cypriniformes. Based on coding sequences （CDS） of tyro- sine kinase regions of IRa and IRb, molecular evolution and phylogenetic relationships were analyzed to better understand the characteristics of IR gene divergence in the family Cyprinidae. 1Ra and IRb were clustered into one lineage in the gene tree of the IR gene family, reconstructed using the unweighted pair group method with arithmetic mean （UPGMA）. IRa and IRb have evolved into distinct genes after IR gene duplication in Cyprinidae. For each gene, molecular evolution analyses showed that there was no significant difference among different groups in the reconstructed maximum parsimony （MP） tree of Cyprinidae; IRa and 1Rb have been subjected to similar evolutionary pressure among different lineages. Although the amino acid sequences of IRa and IRb tyrosine kinase regions were highly conserved, our analyses showed that there were clear sequence variations between the tyrosine kinase regions of IRa and IRb proteins. This indicates that IRa and IRb proteins might play different roles in the insulin signaling pathway.

Inhibition of phosphoenolpyruvate carboxykinase gene expression by metformin in cultured hepatocytes

Objective To investigate the effect and mechanism of the antihyperglycemic agent metformin on the expression of phosphoenolpyruvate carboxykinase (PEPCK) gene in hepatocytes and to determine whether the effects of metformin in hepatocytes are transmitted throughout the known insulin signaling pathways Methods Confluent H4IIE rat heptoma cells were cultured for 16 h with 0 1 mmol/L metformin either in absence or presence of 0 1 nmol/L insulin, and then stimulated with various agents The expression of PEPCK gene was examined by Northern blot analysis Results Therapeutic concentrations of metformin significantly inhibited basal PEPCK mRNA expression and also decreased cAMP and dexamethasone induced PEPCK gene expression through interaction with insulin In the presence of insulin signaling pathway inhibitors wortmannin and UO126, metformin reduced PEPCK mRNA levels, but wortmannin blocked inhibitory regulation of insulin on PEPCK gene expression Conclusion Metformin inhibits PEPCK gene expression via either an insulin independent or an interacting with insulin manner The results suggest that a possible mechanism by which metformin reduces gluconeogenesis could be associated with the inhibition of PEPCK gene expression