Phagocytosis Conditions of Peripheral Blood Leucocytes in Chicken

[Objective] To investigate the optimal conditions of peripheral blood leucocytes phagocytosing Staphy/ococcus spp. [Method] With Staphylococcus spp. 10 145 as tested materials, phagocytosis activity of chicken＇s peripheral blood leucocytes was detected by the method of Wright＇s staining. The effects of time, temperature and concentration of Staphylococcus spp. 10 145 on phagocytosis percentages （PP） and phagocytosis index （PI） were optimized by orthogonal test and their morphologic characteristics were observed by microscope. [ Result] The concentration of Staphylococcus spp. 10 145 was the primary factor effecting leucocytes phagocytosis index. The optimal phagocytosis conditions were at the temperature of 41℃, time of 40 min and concentration of Staphylococcus spp. 10 145 of 5 ×10^ 6/ml. The order of various factors on leucocytes phagocytcsis index was the concentration of Staphylococcus spp. 10 145 〉 time 〉 temperature. [ Conclusion] This study provides a more simple and reliable method for detecting leucocytes phagocytosis function.

INFLUENCES OF MOLECULAR WEIGHT AND BRANCHING PARAMETER OF LACQUER POLYSACCHARIDE ON THE GROWTH OF LEUCOCYTES

A method of determining branching parameter of lacquer polysaccharide wasestablished by acid-base back-titration of terminal uronic acid of branches. The branchingfactors obtained are in agreement with the values determined by colorimetric method withcarbazole and the results estimated by using Zimm-Stockmayer equation from viscositydata. Influences of molecular weights and branching factors of five fractions of lacquerpolysaccharide on the bioactivities were studied. The results show that the polysaccharideshave bioactivities in motivating the growth of leucocytes, and the effect increases with thedecrease of molecular weight and branching factor in the range studied (17×10~4 >M_w>4×10~4).

In vitro immunotoxicity of untreated and treated urbanwastewaters using various treatment processes to rainbow trout leucocytes

Municipal effluents are known to impede the immune system of aquatic organisms. The purpose of this studywas to examine the immunotoxicity of urbanwastewaters before and after 6 treatment processes from 12 cities toward trout leucocytes. Freshly prepared trout leucocyteswere exposed to increasing concentrations of solid phase （C18） extracts ofwastewaters for24 hr at 15°C. Immunocompetencewasdetermined by following changes in leucocyte viability and the proportion of cells able to ingest at least one （immunoactivity） and at least three （immunoefficiency） fluorescent beads. The influentswere treated by sixdifferent treatment strategies consisting of facultative aerated lagoons, activated sludge, biological aerated filter, biological nutrient removal, chemically-assisted physical treatment and trickling filter/solid contact. Water quality parameters of thewastewaters revealed that the plants effectively removed total suspended solids and reduced the chemical oxygendemand. The results revealed that the effluents＇ immunotoxic propertieswere generally more influenced by the properties of the untreatedwastewaters than by the treatment processes. About half of the incoming influentsdecreased leucocyte viabilitywhile4 treatment plantswere able to reduce toxicity. The influents readily increased phagocytosis activity for 8/12 influentswhile itwasdecreased in4/12 influents. This increasewas abolished for4/12 of the effluents using treatments involving biological and oxidative processes. In conclusion, municipal effluents have the potential to alter the immune system in fish and more researchwill be needed to improve the treatments ofwastewaters to better protect the quality of the aquatic environment.

Effect of Allitridi on Peripheral Leucocytes Deformation of Patients with Acute Cerebral Infarction

Objective: To prove the effect of Allitridi in treating acute cerebral infarction. Methods: Millipore membrane filtering technique was adopted to systematically observe the blood rheology and the peripheralleucocytes filtering index of 53 healthy subjects and that of 33 patients before and after treatments, who sufferedfrom acute cerebral infarction and were confirmatorily diagnosed through CT. Results: The filtering index of peripheral leucocytes during the acute period of cerebral infarction rose significantly (6. 1397±4. 4602), and thedifference was significant compared with that of the healthy subjects (0. 8651±0. 4603, P＜0. 01 ). Treatedwith Allitridi, the patients' symptoms got improved and at the same time the filtering index of leucocytes lowered markedly(1. 6261±1. 3472). The conditions of the patients before and after treatments were obviously different(p＜0. 01). The index of the whole blood rheology was improved significantly. Conclusions: Allitridicould effectively improve the leucocytes deformation and the whole blood rheology as well, and alleviate clinicalsymptoms.

Treating inflammatory bowel disease by adsorptive leucocytapheresis:A desire to treat without drugs

Ulcerative colitis and Crohn’s disease are the major phenotypes of the idiopathic inflammatory bowel disease (IBD), which afflicts millions of individuals throughout the world with debilitating symptoms, impairing function and quality of life. Current medications are aimed at reducing the symptoms or suppressing exacerbations. However, patients require life-long medications, and this can lead to drug dependency, loss of response together with adverse side effects. Indeed, drug side effects become additional morbidity factor in many patients on long-term medications. Nonetheless, the efficacy of anti-tumour necrosis factors (TNF)-α biologics has validated the role of inflammatory cytokines notably TNF-α in the exacerbation of IBD. However, inflammatory cytokines are released by patients’ own cellular elements including myeloid lineage leucocytes, which in patients with IBD are elevated with activation behaviour and prolonged survival. Accordingly, these leucocytes appear logical targets of therapy and can be depleted by adsorptive granulocyte/monocyte apheresis (GMA) with an Adacolumn. Based on this background, recently GMA has been applied to treat patients with IBD in Japan and in the European Union countries. Efficacy rates have been impressive as well as disappointing. In fact the clinical response to GMA seems to define the patients’ disease course, response to medications, duration of active disease, and severity at entry. The best responders have been first episode cases (up to 100%) followed by steroid naïve and patients with a short duration of active disease prior to GMA. Patients with deep ulcers together with extensive loss of the mucosal tissue and cases with a long duration of IBD refractory to existing medications are not likely to benefit from GMA. It is clinically interesting that patients who respond to GMA have a good long-term disease course by avoiding drugs including corticosteroids in the early stage of their IBD. Additionally, GMA is very much favoured by patients for its good safety profile. GMA in 21st century reminds us of phlebotomy as a major medical practice at the time of Hippocrates. However, in patients with IBD, there is a scope for removing from the body the sources of pro-inflammatory cytokines and achieve disease remission. The bottom line is that by introducing GMA at an early stage following the onset of IBD or before patients develop extensive mucosal damage and become refractory to medications, many patients should respond to GMA and avoid pharmacologics. This should fulfill the desire to treat without drugs.

Efficacy of boswellic acid on lysosomal acid hydrolases,lipid peroxidation and anti-oxidant status in gouty arthritic mice

Objective:To evaluate the efficacy of boswellic acid against monosodium urate crystal-induced inflammation in mice.Methods:The mice were divided into four experimental groups.GroupⅠserved as control;mice in groupⅡwere injected with monosodium urate crystal;groupⅢconsisted of monosodium urate crystal-induced mice who were treated with boswellic acid(30mg/kg/b.w.);groupⅣcomprised monosodium urate crystal-induced mice who were treated with indomethacin(3mg/kg/b.w.).Paw volume and levels/activities of lysosomal enzymes,lipid peroxidation,anti-oxidant status and inflammatory mediator TNF-αwere determined in control and monosodium urate crystal-induced mice.In addition,the levels ofβ-glucuronidase and lactate dehydrogenase were also measured in monosodium urate crystal-incubated polymorphonuclear leucocytes(PMNL)in vitro.Results:The activities of lysosomal enzymes,lipid peroxidation,and tumour necrosis factor-αlevels and paw volume were increased significantly in monosodium urate crystal-induced mice,whereas the activities of antioxidant status were in turn decreased.However,these changes were modulated to near normal levels upon boswellic acid administration.In vitro,boswellic acid reduced the level ofβ-glucuronidase and lactate dehydrogenase in monosodium urate crystal-incubated PMNL in concentration dependent manner when compared with control cells.Conclusions:The results obtained in this study further strengthen the anti-inflammatory/antiarthritic effect of boswellic acid,which was already well established by several investigators.

ABO血型不合肾移植的现状及研究进展

肾移植是治疗终末期肾病最有效的方法。供体来源严重短缺是限制器官移植发展的一大瓶颈。积极开展器官捐献和亲属活体肾移植,是解决供肾短缺的重要途径（[1-2]）。随着围术期和术后管理方案的不断进步,肾移植逐渐突破了人类白细胞抗原（human leucocyte antigen,HLA）配型和ABO血型的限制（[3]）。据统计,潜在亲属活体肾移植中,

尾静脉注射和鼻腔接种禽流感H5N1亚型病毒对血细胞的影响

[目的]Balb/c小鼠通过尾静脉注射和鼻腔接种禽流感H5N1亚型病毒造疾病模型,观察其血细胞变化。[方法]以禽流感H5N1亚型病毒通过静脉和鼻腔接种Balb/c小鼠,分别在0~7天和0~11天取血进行血细胞计数和分类,同时对于静脉接种方法进行了0~8小时的短时间监控。[结果]静脉接种途径在接种病毒后2小时就能显著的降低小鼠的白细胞总数、粒细胞及淋巴细胞数,白细胞总数及淋巴细胞数在第3天、粒细胞数在第2天就恢复正常而后均显著升高;而鼻腔接种仅在1~3天淋巴细胞有所降低,而后恢复正常;1~6天白细胞总数及粒细胞数显著升高,而后恢复正常。[结论]从血细胞变化而言,静脉接种相比鼻腔接种方法要好。

Studies on the mechanisms of anti-inflammatory activity of the extracts and fractions of Alchornea floribunda leaves

Objective:Alchornea floribunda leaves are widely used in ethnomedicinal management of inflammatory disorders. The present work is aimed at investigating this folkloric use.Methods:The anti-inflammatory effect of the leaf extracts and fractions was investigated in experimental animal models of acute and chronic inflammation. The possible mechanisms by which the two most active fractions,hexane(HE) and ethyl acetate(EF) exert their effects were also investigated.Results:The crude extract(200 mg/kg) showed moderate inhibition of egg albumen-induced edema in rats(%edema inhibition = 54.69) at 4 h.HE and EF showed very high activity(%edema inhibition of 81.25 and 67.19 respectively at 200 mg/kg) at 4h as compared to the control. Both fractions ameliorated arthritis induced by formaldehyde in rats.At 400 mg/kg,HE evofeed a significant irritation of gastric mucosa in rats.EF(200 mg/kg,p.o.) significantly inhibited leucocytes(%inhibition = 36.79) migration in vivo,but could not stabilize heat and hypotonicity-indueed lysis of human erythrocyte at 200 and 400μg/mL in vitro.Phytochemical investigation revealed the presence of terpenoids and steroids in HE and flavonoids,tannins and saponins in EF.Conclusion:These results suggest that the leaves of Alchornea floribunda possess anti-inflammatory activity in acute and chronic inflammation.The activity may derive from a combination of inhibition of prostaglandin synthesis and leucocytes migration.The phytochemical constituents detected in HE and EF may account for the anti-inflammatory activity.

Sishengtang decoction in alleviation of toxic and side effects of transarterial embolization

AIM To observe the therapeutic effects of Sishengtang decoction in alleviating the toxic and side effects of transarterial embolization (TAE). METHODS Fifty four patients with liver cancer were divided randomly into Sishengtang decoction group (34 cases) and control group (20 cases). The changes of clinical symptoms and peripheral hemogram and some cellular immune functions were observed before and two weeks after TAE. RESULTS Sishengtang decoction was superior to the control group in improving the digestive tract reaction. The leucocytes of peripheral blood and cellular immune functions (activities of NK cells and LAK cells) of control group decreased obviously after TAE, while that of Sishengtang decoction group decreased slightly, without obvious difference as compared with that of preoperation. CONCLUSIONS Sishengtang decoction might improve the clinical symptoms and increase the leucocytes of peripheral blood and the cellular immune functions of TAE patients.

遗传性共济失调的遗传及生化研究

文献报道遗传性共济失调已有60多种类型,通常为家族遗传性。近年研究发现本病可能有HLA 相关和非相关两型,并发现病人的神经递质和酶在量及代谢方面都发生改变。把症状学、病理发现和脑尸检的生化改变结合起来,将可进一步了解本病的异型和分类,并对患者的预后作出更准确的估计。

Spontaneous bacterial peritonitis

Since its initial description in 1964,research hastransformed spontaneous bacterial peritonitis (SBP)from a feared disease (with reported mortality of 90%)to a treatable complication of decompensated cirrhosis,albeit with steady prevalence and a high recurrencerate. Bacterial translocation,the key mechanism in thepathogenesis of SBP,is only possible because of theconcurrent failure of defensive mechanisms in cirrhosis.Variants of SBP should be treated. Leucocyte esterasereagent strips have managed to shorten the 'tap-to-shot' time,while future studies should look into theircombined use with ascitic fluid pH. Third generationcephalosporins are the antibiotic of choice becausethey have a number of advantages. Renal dysfunctionhas been shown to be an independent predictor ofmortality in patients with SBP. Albumin is felt to reducethe risk of renal impairment by improving effectiveintravascular volume,and by helping to bind pro-inflammatory molecules. Following a single episodeof SBP,patients should have long-term antibioticprophylaxis and be considered for liver transplantation.

Genetics of inflammatory bowel disease: The role of the HLA complex

The human leucocyte antigen （HLA） complex on chromosome 6p21.3 is the most extensively studied genetic region in Inflammatory bowel disease （IBD）. Consistent evidence of linkage to IBD3 （6p21.1-23）, an area which encompasses the HLA complex, has been demonstrated for both Crohn＇s disease and ulcerative colitis, and a number of replicated associations with disease susceptibility and phenotype have recently emerged. However, despite these efforts the HLA susceptibility gene （s） for IBD remain elusive, a consequence of strong linkage disequilibrium, extensive polymorphism and high gene density across this region. This article reviews current knowledge of the role of HLA complex genes in IBD susceptibility and phenotype, and discusses the factors currently limiting the translation of this knowledge to clinical practice.

Genetic association of autoimmune hepatitis and human leucocyte antigen in German patients

AIM: To report on our large German collective and updated data of 142 patients with autoimmune hepatitis (AIH) type 1. METHODS: Key investigations performed were liver biopsy, serum autoantibodies as well as serum markers such as IgG and elevated transaminases. Antinuclear antigen (ANA) and smooth muscle antigen (SMA) autoantibodies characterized type 1 AIH. Type 3 (AIH) was solely characterized by the occurrence of soluble liver antigen/liver-pancreas antigen (SLA/LP) autoantibodies either with or without ANA or SMA autoantibodies. RESULTS: Most prevalent HLAs were A2 (68 patients, 48%), B8 (63 patients, 44%), C7 (90 patients, 63%), DR3 (49 patients, 38%), DR4 (49 patients, 38%) and DQ2 (42 patients, 30%). Compared to the Italian and North American patients, we found fewer patients with a DQ2 subtype. Furthermore, the B8-DR3-DQ2 human leucocyte antigen (HLA) was also less prominent compared to the North American patients. However, prevalences of B8, DR3, DR4, DR7, DR11 and DR13 were comparable to the Italian and North American patients. Furthermore, we report on an additional subgroup of patients with SLA/LP positive AIH. Generally, in this subgroup of patients the same HLA subtypes were favoured as the AIH type 1. CONCLUSION: Although HLA subtypes were comparable between these three collectives, the German patients were distinct from the Italian and North American patients with respect to DQ2 and from the North American patients with respect to B8-DR3-DQ2HLA. A clinical correlation, e.g. difference in severity or treatability of AIH type 1, has yet to be determined.

The human leucocyte differentiation antigens (HLDA) workshops: the evolv-ing role of antibodies in research, diagnosis and therapy

The 8^th International Workshop on Human Leucocyte Differentiation Antigens （chaired by HZ and managed by BS） was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievements of the HLDA Workshops and provide links to information on CD molecules and antibodies against them, including the 93 new CDs assigned in the 8^th Workshop. We consider what remains to be achieved （including an estimate of the number of leucocyte surface molecules still to be discovered）, and how the field can best move forward.

HLA-DQ:Celiac disease vs inflammatory bowel disease

AIM To determine the genetic predisposition to celiacdisease(Ce D) in inflammatory bowel disease(IBD) patients by quantifying the frequency of Ce D-related human leucocyte antigen(HLA)(HLA-Ce D: HLA-DQ2 and-DQ8) in IBD patients globally, by type of IBD and gender, and by calculating the protective/risk contribution of these haplotypes in the development of the IBD disease.METHODS We conducted a prospective study with IBD patients from our Unit. Clinical information was gathered and blood was tested for HLA-CeD. The control group was made up of unrelated Valencian organ donors.RESULTS1034 subjects were analyzed: 457 IBD [207 ulcerative coliti(UC) and 250 Crohn's disease(CD)] patients and 577 healthy controls. 39% of the controls and 34% of the patients had HLA-Ce D(P = 0.0852). HLA-DQ2 was less frequent in UC patients(P = 0.0287), and HLA-DQ8 in CD(P = 0.0217). In women with UC, the frequency of DQ2.5 cis(DQB1*02:01-DQA1*05:01) was reduced ≥ 50% [P = 0.0344; preventive fraction(PF) = 13%]. PFs(7%-14%) were obtained with all HLACe D haplotypes. HLA DQB1*02:02-DQA1*02:01(HLADQ2.2) was more frequent in CD patients with respect to controls(P = 0.001) and UC patients(etiological fraction = 15%).CONCLUSION HLA-CeD is not more frequent in IBD patients, with an even lower frequency of HLA-DQ2 and-DQ8 in UC and CD respectively. HLA-DQ2.5 confers protection from the development of UC, especially in women, and HLADQ8 does so for the appearance of CD. HLA-DQ2.2 is present in 34% of the CD patients and may constitute a genetic risk factor for CD development.

Study of Swine Leukocyte Antigen Class I-3 (SLA-3) Gene for Inbreeding Wuzhishan Pig

To elucidate the structure of SLA-3 alleles on inbred line of Wuzhishan pig （WZSP） population, we examined the partial exon 1, completed exon 2, and partial exon 3 of SLA-3 loci using the reverse transcription-polymerase chain reaction （RT- PCR） and the sequencing-based method in 32 WZSPs. According to pedigree and amplification results, PCR products of 8 WZSPs were selected to clone and sequence. Nine different nucleotide sequences were obtained. After comparing the DNA and protein sequences of the WZSPs SLA-3 alleles with the published GenBank SLA sequences, it was found that the SLA-3 alleles in WZSPs were all novel, but there were very few variations among them. Comparision of SLA-3 and HLA-A protein sequences indicated that there was more sequence homology. Meanwhile, the construction of a phylogenetic tree using the nucleotide sequences of 23 SLA-3 alleles and 1 HLA-A allele represented that the WZSP population owns its unique genetics resource. In this study, the alleles of SLA-3 on WZSP group were successfully detected and analyzed, which provided the firm basis on the genotype of SLA-3 for breeding specific haplotypes WZSPs.

Axonal growth inhibitors and their receptors in spinal cord injury:from biology to clinical translation

Axonal growth inhibitors are released during traumatic injuries to the adult mammalian central nervous system, including after spinal cord injury. These molecules accumulate at the injury site and form a highly inhibitory environment for axonal regeneration. Among these inhibitory molecules, myelinassociated inhibitors, including neurite outgrowth inhibitor A, oligodendrocyte myelin glycoprotein, myelin-associated glycoprotein, chondroitin sulfate proteoglycans and repulsive guidance molecule A are of particular importance. Due to their inhibitory nature, they represent exciting molecular targets to study axonal inhibition and regeneration after central injuries. These molecules are mainly produced by neurons, oligodendrocytes, and astrocytes within the scar and in its immediate vicinity. They exert their effects by binding to specific receptors, localized in the membranes of neurons. Receptors for these inhibitory cues include Nogo receptor 1, leucine-rich repeat, and Ig domain containing 1 and p75 neurotrophin receptor/tumor necrosis factor receptor superfamily member 19(that form a receptor complex that binds all myelin-associated inhibitors), and also paired immunoglobulin-like receptor B. Chondroitin sulfate proteoglycans and repulsive guidance molecule A bind to Nogo receptor 1, Nogo receptor 3, receptor protein tyrosine phosphatase σ and leucocyte common antigen related phosphatase, and neogenin, respectively. Once activated, these receptors initiate downstream signaling pathways, the most common amongst them being the Rho A/ROCK signaling pathway. These signaling cascades result in actin depolymerization, neurite outgrowth inhibition, and failure to regenerate after spinal cord injury. Currently, there are no approved pharmacological treatments to overcome spinal cord injuries other than physical rehabilitation and management of the array of symptoms brought on by spinal cord injuries. However, several novel therapies aiming to modulate these inhibitory proteins and/or their receptors are under investigation in ongoing clinical trials. Investigation has also been demonstrating that combinatorial therapies of growth inhibitors with other therapies, such as growth factors or stem-cell therapies, produce stronger results and their potential application in the clinics opens new venues in spinal cord injury treatment.

Asymptomatic bacteriuria in pregnancy in Port Harcourt

Objective:To determine the prevalence of asymptomatic bacteriuria in pregnancy,to consider the antimicrobial sensitivity patterns of involved uropathogens,to elucidate the safety profiles of antibacterial agents,and to evaluate the role of urinalysis in screening for asymptomatic bacteriuria.Methods:About 760 apparently healthy pregnant subjects attending the Antenatal Clinic of the University of Port Harcourt Teaching Hospital were randomly selected for this study.Urinalysis and microscopy,culture,and sensitivity tests were carried out on clean-catch midstream urine samples obtained from subjects.Biochemical reagent strips were used for urinalysis while the standard wire loop and agar diffusion technique were respectively employed for culture and susceptibility testing.Results:A total of 111 samples yielded moderate or severe growth on culture after 48 hours comprising 35,31,27,and 18 isolates of Staphylococcus spp.,Proteus spp.,Klebsiella spp.,and Escherichia spp,respectivehy.Urinalysis results were positive for the presence of nitrate reductase and leucocyte esterase activity in 17 urine samples of these 111 samples.The isolates showed a general sensitivity to the fluorinated quinolones and to Nitrofurantoin.Conclusions:The prevalence of asymptomatic bacteriuria is 14.6%, with the predominant organism being Staphylococcus spp.Drugs used for treatment should have excellent fetal safety profiles,and a rapid screening test with a high negative predictive value for asymptomatic bacteriuria would be ideal.