Effects of obstructive jaundice on neutrophil production and acquisition of chemotactic activity in the bone marrow

BACKGROUND AND AIMS: Increased numbers and enhanced functions of peripheral neutrophils have been observed in obstructive jaundice. However, the effects of obstructive jaundice on the bone marrow, that is neutrophil production and acquisition of neutrophil chemotactic activity, have been poorly understood. In the present study, differentials of bone marrow cells and chemotactic activity of bone marrow neutrophils were evaluated in bile duct-obstructed rats. METHODS: Male Wistar rats underwent either bile duct obstruction for 10 days or bile duct obstruction for 4 days followed by 6 days' internal biliary drainage. Differentials of peripheral blood and bone marrow cells were sequentially determined. Chemotactic activity of peripheral and bone marrow neutrophils was evaluated with a modified Boyden method using interleukin-8 (recombinant rat Gro-beta) as a chemoattractant. RESULTS: Numbers of peripheral neutrophils significantly increased after bile duct obstruction. Significant increases in the myeloid/erythroid (M/E) ratio of bone marrow cells were observed after bile duct obstruction. The neutrophil proliferative pool (promyelocytes and myelocytes) increased initially, followed by an increased neutrophil storage pool (metamyelocytes, bands, and segmented neutrophils). The M/E ratio as well as the neutrophil proliferative and storage pools normalized after internal biliary drainage. Chemotactic activity was enhanced in both peripheral and bone marrow neutrophils after bile duct obstruction, and enhanced chemotaxis was alleviated with internal biliary drainage. CONCLUSION: The present results strongly suggest the principal role of the bone marrow in increasing the number of neutrophils and their chemotactic activity during obstructive jaundice.     

Effect of obstructive jaundice on neutrophil chemotactic activity: An in vivo assessment in zymosan-induced peritonitis model in rats

The effect of obstructive jaundice on local neutrophil accumulation in response to inflammatory stimulus was investigated in rats. Obstructive jaundice was produced by bile duct ligation for 7 days. Zymosan (200 mg) was injected intraperitoneally and 4h later myeloperoxidase activity in the peritoneal fluid was measured to quantify neutrophil recruitment. Zymosan-induced neutrophil recruitment was significantly greater (more than two-fold) in bile duct-ligated rats than in sham-ligated or normal animals. Depletion of peritoneal cells significantly suppressed neutrophil recruitment after zymosan injection in all three groups, with no significant differences between the groups. In normal rats, replacement of their peritoneal cells by those from bile duct-ligated rats did not enhance zymosan-induced neutrophil recruitment. In contrast, bile duct-ligated rats treated with peritoneal cell replacement from normals showed significantly increased neutrophil recruitment after zymosan injection. In vitro neutrophil chemotaxis in response to formyl-Met-Leu-Phe was significantly enhanced in bile duct-ligated rats, compared with that in sham-ligated animals. The results suggest that local neutrophil recruitment in response to inflammation may be enhanced in obstructive jaundice and that increased neutrophil chemotactic activity, not macrophage activity, may play a prime role in the mechanism.     

Increased neutrophil chemotaxis in obstructive jaundice: An in vitro experiment in rats

BACKGROUND AND AIMS: Changes in neutrophil functions in obstructive jaundice have been poorly understood. An in vitro experimental study was performed to evaluate the effect of obstructive jaundice on the functions of macrophages (secretion of neutrophil chemoattractants) and neutrophils (chemotaxis and superoxide anion generation). METHODS: Obstructive jaundice was produced in rats by 7 day bile duct ligation. Peripheral neutrophils and peritoneal macrophages were harvested from either normal, sham-ligated or bile duct-ligated rats and supernatants of the monolayers of the respective macrophages were prepared after stimulation with lipopolysaccharide. Neutrophil chemotaxis was evaluated with a modified Boyden method. RESULTS: The supernatant of the bile duct-ligated rat macrophages showed a chemotactic effect on normal rat neutrophils with insignificant difference from the supernatant of the sham-ligated rat macrophages. Chemotaxis of the bile duct-ligated rat neutrophils towards the supernatant of the normal rat macrophages was significantly increased, compared with that of sham-ligated rat neutrophils. Similarly, neutrophils from bile duct-ligated rats showed significantly greater chemotaxis to formyl-methionyl-leucyl-phenylalanine than the sham-ligated rat neutrophils. Superoxide anion generation in response to formyl-methionyl-leucyl-phenylalanine or phorbol myristate acetate was significantly increased in the bile duct-ligated rats compared with the sham-ligated rats. CONCLUSIONS: The results suggest that the neutrophil is primed in terms of chemotaxis and superoxide anion generation in obstructive jaundice. How these activated neutrophils play a role in the inflammatory response to obstructive jaundice should be evaluated.     

108例阻塞性黄疸的超声诊断分析

目的 探讨超声在阻塞性黄疸诊断中的价值.方法 对我院2004-2007年收治并手术的108例阻塞性黄疸患者的超声声像图进行回顾性分析和总结.结果 阻塞性黄疸超声诊断与手术病理结果符合102例(94.4%),其中胆管结石65例,诊断符合率100%;胆管狭窄2例,诊断符合率66.6%;胰腺炎1例,诊断符合率50.0%;胆囊炎(向后压迫)3例,诊断符合率100%;胰头癌14例,诊断符合率87.5%;壶腹癌11例,诊断符合率91.7%;胆管癌6例,诊断符合率85.7%.结论 超声诊断阻塞性黄疸与手术病理结果符合率高,可作为阻塞性黄疸检查的首选方法,结合临床可提高诊断率.     

覆膜支架治疗梗阻性黄疸的价值分析

[目的]探讨覆膜支架在治疗梗阻性黄疸患者中的临床应用价值。[方法]选择梗阻性黄疸行胆道覆膜支架置入患者30例,随访1年,对其术后产生的并发症进行分析。[结果]30例手术成功率、有效率100%,胆道感染发生率10.0%,胆道出血发生率6.7%,胆囊炎发生率3.3%,胰腺炎发生率6.7%,支架滑脱、移位发生率10.0%,支架阻塞发生率6.7%。[结论]覆膜支架可以有效防止胆道再狭窄或闭塞;其滑脱、移位发生率较高;不易跨越胆囊管开口放置,可以跨越壶腹放置。     

Pancreatic enlargement in obstructive jaundice

To verify the influence of obstructive jaundice on pancreatic growth, the anteroposterior width of the pancreas was measured by computed tomography in 30 cholangiocarcinoma patients excluded patients with distal bile duct tumor (jaundice group) and 74 control subjects. Follow-up examinations were performed on 12 patients with and without internal biliary drainage to elucidate the temporal relationship between pancreatic enlargement and the diversion of the obstructed biliary stream. Histologic analysis on autopsy samples from 13 control and 10 jaundice cases also was performed. Mean pancreatic head and body widths in the jaundice group were 2.93±0.3 cm and 2.01±0.3 cm, respectively. These values were significantly greater than those of the controls (2.13±0.3 cm and 1.49±0.3 cm,P<0.01). The glandular widths returned to their normal sizes following internal biliary drainage. No changes were seen in patients who underwent external drainage alone. Histologic examination revealed that enlargement of the acinar cells or of the islet of Langerhans was often seen in the jaundiced patients. Therefore obstructive jaundice is thought to cause pancreatic growth through a trophic effect by interrupting biliary circulation.     

Cytokine production by bone marrow mononuclear cells in inherited bone marrow failure syndromes

Fanconi anaemia (FA), dyskeratosis congenita (DC), Diamond‐Blackfan anaemia (DBA), and Shwachman‐Diamond syndrome (SDS) are characterized by the progressive development of bone marrow failure. Overproduction of tumour necrosis factor‐α (TNF‐α) from activated bone marrow T‐cells has been proposed as a mechanism of FA‐related aplasia. Whether such overproduction occurs in the other syndromes is unknown. We conducted a comparative study on bone marrow mononuclear cells to examine the cellular subset composition and cytokine production. We found lower proportions of haematopoietic stem cells in FA, DC, and SDS, and a lower proportion of monocytes in FA, DC, and DBA compared with controls. The T‐ and B‐lymphocyte proportions were similar to controls, except for low B‐cells in DC. We did not observe overproduction of TNF‐α or IFN‐γ by T‐cells in any patients. Induction levels of TNF‐α, interleukin (IL)‐6, IL‐1β, IL‐10, granulocyte colony‐stimulating factor, and granulocyte‐macrophage colony‐stimulating factor in monocytes stimulated with high‐dose lipopolysaccharide (LPS) were similar at 4 h but lower at 24 h when compared to controls. Unexpectedly, patient samples showed a trend toward higher cytokine level in response to low‐dose (0·001 μg/ml) LPS. Increased sensitivity to LPS may have clinical implications and could contribute to the development of pancytopenia by creating a chronic subclinical inflammatory micro‐environment in the bone marrow.     

Increased renal susceptibility to gentamicin in the rat with obstructive jaundice

To study whether renal susceptibility to nephrotoxic stimuli is increased in obstructive jaundice, the effect of gentamicin on the renal function in bile duct-ligated rats was investigated. Gentamicin (50 mg/kg/day, subcutaneously) or saline was given to bile duct-ligated rats or sham-operated rats for six days. Mortality in the bile duct-ligated group that received gentamicin was 64% whereas that in the other groups was 0%. In the bile duct-ligated group, although serum creatinine and blood urea nitrogen were minimally affected, focal granulovacuolar degeneration in the proximal tubule was observed, which was accompanied by an increase in renal malondialdehyde. Gentamicin significantly increased serum creatinine and blood urea nitrogen levels and caused marked degeneration in the proximal tubule in the bile duct-ligated group, which was accompanied by a further increase in renal malondialdehyde, while these changes were not observed in the sham group. The kidney in obstructive jaundice appears to be susceptible to gentamicin. Enhanced production of oxygen radicals may be responsible for this effect.     

Three cases of small hepatocellular carcinoma presenting as obstructive jaundice

Background

Despite improved diagnostic tools, it is often difficult to make a correct diagnosis of small hepatocellular carcinoma (HCC) in patients with obstructive jaundice.

Case outlines

Three cases of small HCC (<2 cm diameter) presenting as obstructive jaundice are reported. All tumours were initially diagnosed as hilar cholangiocarcinoma based on ultrasono-graphy, computed tomography, cholangiography and angiogra-phy. Because of insufficient hepatic function, none of the patients underwent hepatic resection. One patient died 8 months after first admission to our hospital, another died of disseminated intravascular coagulation I month after admission, and the third was treated with hepatic arterial infusion chemotherapy and survived >36 months.

Conclusion

It is important to consider HCC in the diagnosis of obstructive jaundice in patients who are predisposed to HCC because of liver cirrhosis and/or chronic viral hepatitis, and have elevated serum alpha-fetoprotein.     

Alterations of tumor necrosis factor-alpha and interleukin 6 production and activity of the reticuloendothelial system in experimental obstructive jaundice in rats

BackgroundImmunological changes are well recognised in obstructive jaundice. The aim of this study was to monitor plasma tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) levels in rats with obstructive jaundice.MethodsThe ability of splenocytes and peritoneal exudate cells (PEC) to produce these cytokines both spontaneously and on induction with lipopolysaccharide (LPS), was compared in rats with and without obstructive jaundice (OJ). The activity of the reticuloendothelial system (RES) was also measured.ResultsSerum cytokine levels in OJ rats were higher than in control rats. PEC cultures produced significantly more IL-6, compared with control rats, declining thereafter. TNF-α activity in the splenocyte cultures of OJ rats was also higher than in the control group. Pronounced differences were found in the ability to produce TNF-α by PEC, i.e., TNF-α production was much stronger on day 7 in OJ rats than in controls. On day 14 TNF-α production was much lower and the spontaneous response was equal to the LPS-induced one. On day 21 the cells of OJ rats partially regained the ability to produce TNF-α RES activity of OJ rats was significantly suppressed in the liver and spleen, whereas the phagocytic activity in the lungs was elevated.ConclusionWe have demonstrated that the immune reactivity of OJ rats, intially elevated, underwent subsequent depression. The study also revealed a major effect of the operation alone on the studied parameters.     

梗阻性黄疸与肠源性内毒素血症的临床研究

以梗阻性黄疸 (OJ)和胆囊结石患者为实验组和对照组 ,探讨梗阻性黄疸是否影响消化道粘膜屏障功能造成肠源性内毒素血症。分光光度法监测外周血中二胺氧化酶 (DAO)和内毒素 (ET)的活性。与正常组比较 ,术前梗阻性黄疸组的DAO活性明显升高 (t=3.176 ,P <0 0 0 1) ;而术前胆囊结石组与正常人DAO活性差异无显著性(t=0 .0 0 77,P >0 0 5 )。梗黄组术前DAO活性明显高于术后 (t=3.86 2 3,P =0 .0 114 )。梗黄组和胆囊结石组术前、术后的DAO、ET活性分别比较 ,差异有显著性 ,但梗黄组的ET含量术后较术前升高 (P <0 0 5 )。相关分析表明梗黄组术前的DAO与ET相关 ,相关系数为r=0 .74 7,P <0 0 1。梗阻性黄疸所致的消化道粘膜屏障功能异常可能与肠源性内毒素血症密切相关。     

Effect of obstructive jaundice on hemodynamics in the liver and its clinical significance

BACKGROUND:Many diseases can cause obstructive jaundice and then lead to a series of pathologic disorders. Thus preoperative assessment of liver function is of utmost importance.Traditional assessment is to monitor related indicators of liver function,but it is invasive and needs to be performed repeatedly.Color Doppler flow imaging (CDFI)was used to monitor blood flow of the hepatic artery and portal vein,a non-invasive method which can be used repeatedly. METHODS:Twenty cases of obstructive jaundice were ...     

Preoperative biliary drainage (stenting) for treatment of obstructive jaundice

The role of preoperative biliary drainage in malignant obstructive jaundice has been controversial. Laboratory studies suggest that relief of jaundice prior to major pancreatic resection would be associated with improved morbidity and mortality. However, clinical experience has not supported the laboratory results. Obstructive jaundice can be relieved preoperatively via an endoprosthesis introduced either percutaneously or endoscopically. Cohort studies have not shown any clinical benefit and in some the endoprostheses have been implicated in postoperative complications. The only randomized study has shown no benefit in preoperative drainage, but one recent study has confirmed that endoscopic drainage, whilst not conferring an advantage, did no harm. Hence, whilst preoperative drainage is not recommended, if for any reason operation needs to be delayed, endoscopic drainage via an endoprosthesis can be used without fear of adversely influencing the outcome.     

Human polymorphonuclear leukocytes of the bone marrow, circulation, and marginated pool: function and granule protein content

Polymorphonuclear leukocytes (PMN) demonstrate altered function during acute infections and after administration of corticosteroids. We questioned whether or not such changes are due to population shifts from functionally different compartments of the granulocyte pool. Volunteers were given epinephrine to induce demargination or hydrocortisone (HC) to promote egress of PMN from the bone marrow. PMN obtained before and after drug administration were compared for adherence, chemotaxis, luminol-enhanced chemiluminescence, and total content and release of lactoferrin (LF), myeloperoxidase (MPO), and beta-glucuronidase (beta-glu). Epinephrine induced a significant neutrophilia of mature PMN (segmented neutrophils), but there were no changes in function or granule protein content. HC induced a significant neutrophilia with segmented neutrophils and immature PMN (bands). Circulating PMN obtained 4 hr after HC administration demonstrated less adherence, increased chemiluminescence, increased MPO release, and decreased MPO content. Band neutrophils, however, were more adherent than segmented PMN and showed a similar decrease in adherence following HC in vivo. Thus alteration of PMN adherence following intravenous corticosteroids is not due to an influx of immature neutrophils. On the other hand, it is possible that MPO content and release and capacity for oxidative metabolism change as PMN mature.     

Effect of L-arginine on calcium in hepatic mitochondrion in rats with obstructive jaundice

BACKGROUND: There is much debate over the regulation of mitochondrial calcium overload and reducing the impairment of energy metabolism in hepatic cells. It has not been reported whether L-arginine (L-Arg) can affect hepatic mitochondrial calcium overload. This study was undertaken to investigate the protective effect of L-Arg on Ca2+ handling of hepatic mitochondrion in rats with obstructive jaundice and to clarify its possible mechanism. METHODS: Seventy-two male SD rats were randomly divided into 3 groups: sham operation+normal saline group (SO group), common bile duct ligation+normal saline group (BDL group), and common bile duct ligation+ L-Arg group (L-Arg group). The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and Ca2+ in rat hepatic mitochondrion were examined at the 7th, 14th and 21st day after operation. RESULTS: The Ca2+ and MDA levels of hepatic mitochondrion increased significantly but their SOD content decreased markedly at each time point in the BDL group. Except at the 21st day, the Ca2+ and MDA, contents of hepatic mitochondrion were significantly lower, and SOD concentrations were higher in the L-Arg group than those in the BDL group at the 7th and 14th day (P<0.01). CONCLUSION: L-Arg has a protective effect on mitochondrion in the early and mid stages of obstructive jaundice.     

Cysteinyl leukotrienes in the bile of patients with obstructive jaundice

Background: Cysteinyl leukotrienes (LTs) are potent proinflammatory mediators. They are predominantly excreted from blood by hepatobiliary elimination. To explore the clinical significance of biliary cysteinyl LTs, we determined their concentration changes in bile during treatment in patients with obstructive jaundice. Methods: Bile samples were obtained during endoscopic or transhepatic biliary drainage. Leukotrienes C₄, D₄, and E₄ were quantified by two-step reversed-phase high-performance liquid chromatography and subsequent radioimmunoassay. Results: The increased excretion of cysteinyl LTs (LTC₄ + LTD₄ + LTE₄) decreased between day 1 and 14 after drainage (means, 171 pmol/h to 79 pmol/h; P < 0.02). During drainage, the excretion was higher when there was additional cholangitis (mean, 225 and 86 pmol/h, with and without cholangitis, respectively; P < 0.001). The concentrations of LTD₄ and LTE₄ were also higher with additional cholangitis than without (LTD₄, mean 6.0 vs 2.0 nM; P < 0.05; LTE₄, 6.8 vs 2.4 nM; P < 0.02, respectively). Biliary LTC₄ was detected only in patients with cholangitis. The biliary excretion of cysteinyl LTs was positively correlated with leukocyte concentration (r = 0.68; P < 0.005) and C-reactive protein (r = 0.73; P < 0.005) in blood. Furthermore, only in the absence of cholangitis, the excretion was positively correlated with serum γ-glutamyl transferase (r = 0.76; P < 0.02) and alanine aminotransferase (r = 0.72; P < 0.02). Conclusions: The excretion of biliary cysteinyl LTs increases with the severity of cholestasis and hepatic inflammation in patients with obstructive jaundice. An additional increase of cysteinyl LTs was observed during bacterial cholangitis. The increased biliary excretion of biologically active cysteinyl LTs may contribute to the aggravation of cholestasis and inflammatory reaction in obstructive jaundice. Received: October 1, 2001 / Accepted: March 8, 2002 Acknowledgements. We are indebted to Dr. Gabriele Jedlitschky and Dr. Inka Leier for valuable advice throughout this work. This work was supported by a fellowship awarded to Dr. Masahito Uemura by the Alexander-von-Humboldt Foundation, Bonn, Germany, and in part by the Deutsches Krebsforschungszentrum, Heidelberg, Germany. Reprint requests to: M. Uemura     

Peripheral blood neutrophil morphology reflects bone marrow dysplasia in myelodysplastic syndromes

Dysplastic features of cells in peripheral blood and bone marrow were studied in 51 patients with myelodysplastic syndromes (MDS) to evaluate the significance of the degree of neutrophil granulation (G-score) and the percentage of pelgeroid polymorphs (ppp) in the peripheral blood, as indices of dysplastic changes in the bone marrow. There was a good correlation between peripheral blood and bone marrow findings, both for G-score figures (r = 0.92, P < 0.01) and ppp (r = 0.82, P < 0.01). Significantly lower G-score figures were found among patients with an increased percentage of bone marrow blasts (P < 0.05), while high ppp correlated with the presence of ring sideroblasts, the degree of bone marrow fibrosis, and findings of complex chromosomal abnormalities. Patients with a high degree of bone marrow dysplasia had significantly lower G-score (P < 0.01) and significantly higher ppp (P < 0.05) figures, than those with less pronounced myelodysplasia. In addition, extreme hypogranulation (G-score <150) or very high ppp (≥20%) was generally a sign of bi- and tri-lineage dysplasia in the bone marrow. The results thus show that quantitative estimation of peripheral blood polymorph dysplasia by G-score figures and ppp seems to reflect the total degree of bone marrow dysplasia in MDS and may serve as a complement to bone marrow evaluation when the diagnosis of MDS is difficult. © 1995 Wiley-Liss, Inc.     

Pilot study: effects of G-CSF on neutrophil ex-vivo function post bone marrow transplantation

Summary The effects of human recombinant granulocyte colony stimulating factor (G-CSF) on neutrophil recovery have been studied in patients undergoing bone marrow transplantation for haematological disease. Nine patients (five autografts and four allografts) were studied while receiving daily doses of G-CSF (range 2-20 μg/kg body weight) were compared to eight patients (four autografts and four allografts) who did not receive G-CSF as a control group. In both groups flow cytometry was used to determine neutrophil IgG Fc receptor (FcRII, FcRIII) expression. Phagocytosis and metabolic burst was assessed using IgG opsonized bacteria. The patients' neutrophils were studied prior to conditioning in autografts and donor cells were studied in allografts. Studies were repeated at neutrophil recovery (neutrophil count 1.0 ± 10⁹/1) and ten days post recovery. At recovery FcRII expression was slightly increased in all groups, whereas the number of cells expressing FcRIII was reduced. This reduction was significant in the patients receiving rG-CSF at recovery and post recovery compared to the pretreatment levels. The ability to phagocytose bacteria in cells from patients receiving G-CSF was also reduced. The associated metabolic burst was significantly reduced in the autograft group but not the allograft group receiving rG-CSF. The reduced function and reduced expression of FCRIII suggests the presence of immature neutrophils in patients receiving rG-CSF post BMT.     

The bone marrow aspirate of healthy subjects

Bone marrow aspirates were obtained from the right or left posterior superior iliac spine of 50 healthy volunteers, 30 men and 20 women. Reference ranges were derived for each cell type and for the myeloid : erythroid (M : E) ratio. The M : E ratio and the percentage of neutrophils were significantly higher in the women and the erythroid component significantly lower. All 28 evaluable men and 11/17 evaluable women had storage iron present in more than trace amounts. The percentage of erythroblasts with detectable iron granules varied very widely, from 3% to 69% in those with more than a trace of storage iron. Minor dyserythropoietic features were present in a high percentage of subjects and 19/50 subjects had one or two dysplastic megakaryocytes. Granulocytic dysplasia was not detected.