LPS或全菌体阻断体内炎性单核细胞向树突状细胞(DC)的转化

DC是位于体内淋巴和一些非淋巴组织的专职性抗原提呈细胞(APC)，它在机体免疫系统防御外来病原微生物的过程中起到很重要的作用。体内的迁移DC可以来源于驻留在外周组织中的DC，也可以来源于血液中的单核细胞。研究表明一些细胞因子和内环境的因素会影响单核细胞向巨噬细胞或向DC的分化。由于DC所起的重要抗原提呈作用，当机体受感染后产生迁移DC的能力就直接决定着获得性免疫能力的强弱。通过皮下注射活的或已灭活的细菌或者LPS，诱导了强烈的局部先天免疫应答从而阻断了单核细胞分化生成DC以及向引流淋巴结(DLN)的迁移。表明强烈的先天免疫应答能抑制机体对颗粒性抗原的获得性免疫反应。

正畸加力过程中外周血和脾脏中炎性单核细胞比例和CD226分子表达水平变化

目的:观测小鼠牙齿正畸加力过程中外周血和脾脏中炎性单核细胞比例和CD226分子表达水平变化。方法:选择6~8周龄雌性C57BL小鼠,随机分为加力1d组(n=5)、加力3d组(n=4)、加力7 d组(n=4)和相应时间的对照组(n=3×3)。届时分离小鼠外周血和脾脏进行单个核细胞流式细胞术检测,在体式显微镜下观察牙齿移动情况;取上颌骨标本切片后行HE染色和免疫荧光染色观测破骨细胞生成情况。结果:随着加力时间延长,破骨细胞生成增多;加力1 d时,外周血中炎性单核细胞比例降低;3 d时,外周血和脾脏中炎性单核细胞比例升高,而CD226分子表达水平均降低;7d时,外周血中炎性单核细胞比例较对照组升高但较加力3 d组降低,外周血和脾脏中炎性单核细胞表面CD226分子表达水平降低。结论:正畸加力过程中,小鼠外周血和脾脏中的炎性单核细胞比例和CD226分子表达水平动态变化可能影响牙齿移动过程。

树突状细胞、炎性单核细胞及巨噬细胞在小鼠感染肺孢子菌过程中变化的研究

目的：研究非免疫抑制小鼠感染肺孢子菌（ Pneumocystic murina，P．murina）过程中树突状细胞、炎性单核细胞以及巨噬细胞的变化和表型特征，以探讨上述细胞在肺孢子菌感染时发挥的作用。方法选择6～8周的雄性野生型C57BL／6小鼠构建肺孢子菌感染动物模型，分为实验组和手术对照组，并设空白对照组。肺孢子菌感染后1周、2周、3周和4周，采用TaqMan实时荧光定量PCR法检测肺孢子菌载量；制作病理切片观察肺组织的炎症程度；采用流式细胞术检测两组小鼠肺脏、外周血和骨髓中树突状细胞、炎性单核细胞和巨噬细胞以及脾脏中炎性单核细胞的数量变化，并检测主要组织相容复合物（ major histocompatability complex， MHC）Ⅱ和CX3C趋化因子受体（ CX3C chemokine receptor， CX3CR1）-1，低表达CC趋化因子受体（ CC chemokine receptor， CCR）-2在感染后肺内各细胞群的表达情况。结果实验组小鼠菌载量呈现先升高再降低的趋势，但差异无统计学意义（ P＞0．05）。肺孢子菌感染3周和4周时，肺组织病理改变较重，可见到肺泡结构不清，间质内有炎性细胞浸润以致肺泡间隔增厚。肺孢子菌感染后，肺内CD11 c＋CD11 b＋树突状细胞数量增高，外周血中CD11c＋CD11b＋树突状细胞数量降低，外周血中的炎性单核细胞数量先降低后增高，差异均有统计学意义（P＜0．05）；但肺内的巨噬细胞的数量变化无统计学意义（P＞0．05）。 CD11c＋CD11b＋树突状细胞高表达MHCⅡ、CX3CR1，少量表达CCR2；炎性单核细胞高表达CCR2，部分表达MHCⅡ，低表达CX3CR1；巨噬细胞高表达CX3CR1，低表达MHCⅡ和CCR2。结论非免疫抑制小鼠感染肺孢子菌后，肺内CD11c＋CD11b＋树突状细胞募集增多，高表达MHCⅡ，参与了肺孢子菌感染的免疫应答。

Prediction of severe acute pancreatitis:Current knowledge and novel insights

Acute pancreatitis (AP) is a common and potentially lethal acute inflammatory process with a highly variable clinical course. It is still unclear why some patients progress to organ failure and others do not. Physicians, ability to predict which patients will develop severe disease is limited. Routine clinical and laboratory data and multi-factorial clinical scores measured on admission and during the first 48 h of hospitalization are currently the standards of care used to estimate the magnitude of the inflammatory response to injury. Current literature highlights several common environmental, metabolic and genetic factors that increase the risk of AP development and subsequent adverse sequelae. Several cytokines have been found to play a critical role in the pathogenesis of AP by driving the subsequent inflammatory response, to include tumor necrosis factor-α (TNF-α), Interleukin-1 (IL-1), IL-6 and monocyte chemotactic protein-1 (MCP-1). Large, prospective studies are still needed to address these questions by identifying AP risk factors and serum biomarkers of severe disease.

Nuclear factor kB activity in patients with acute severe cholangitis

AIM: To determine the NF-kB activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NF-kB activation with severity of biliary tract infection and clinical outcome. METHODS: Twenty patients with ACST were divided into survivor group (13 cases) and nonsurvivor group (7 cases). Other ten patients undergoing elective gastrectomy or inguinal hernia repair were selected as control group. Peripheral blood samples were taken 24 hours postoperatively. PBMC were separated by density gradient centrifugation, then nuclear proteins were isolated from PBMC, and Electrophoretic Mobility Shift Assay (EMSA) used determined. The results were quantified by scanning densitometer of a Bio-Image Analysis System and expressed as relative optical density (ROD). The levels of TNF-alpha, IL-6, and IL-10 in the plasma of patients with ACST and healthy control subjects were determined by using an enzyme-linked immunoassay (ELISA). RESULTS: The NF-kB activity was 5.02 +/- 1.03 in nonsurvivor group, 2.98 +/- 0.51 in survivor group and 1.06 +/- 0.34 in control group. There were statistical differences in three groups (P【0.05). The levels of TNF-alpha and IL-6 in plasma were (498 +/- 53)ng.L(-1)and (587 +/- 64)ng.L(-1)in nonsurvivor group, (284 +/- 32)ng.L(-1) and (318 +/- 49)ng.L(-1)in survivor group and (89 +/- 11)ng.L(-1) and (102 +/-13)ng.L(-1)in control group. All patients with ACST had increased levels of TNF-alpha and IL-6, which were many-fold greater than those of control group, and there was an evidence of significantly higher levels in those of nonsurvivor group than that in survivor group (P【0.05). The levels of IL-10 in plasma were (378+/-32)ng.L(-1), (384+/-37)ng.L(-1) and (68+/-11)ng.L(-1) in three groups, respectively. All patients had also increased levels of IL-10 when compared with control group (P【0.05), but the IL-10 levels were not significantly higher in nonsurvivors than in survivors (P】0.05). CONCLUSION: NF-kB activity in PBMC in patients with ACST increases markedly and the degree of NF-kB activation is correlated with severity of biliary tract infection and clinical outcome.

Analysis of monocyte chemotactic protein-1 gene polymorphism in patients with spontaneous bacterial peritonitis

AIM:To investigate a genetic polymorphism of the monocyte chemotactic protein-1 (MCP-1 ) gene in patients with spontaneous bacterial peritonitis (SBP).METHODS:MCP-1 genotyping was performed in 23 patients with SBP and 83 cirrhotic control patients with non-infected ascites.RESULTS:The frequency of carriers of the G-allele was lower in SBP patients but this difference did not reach statistical significance. However,in the subgroup of patients with alcoholic cirrhosis (n=80),carriers of the G-allele were significantly less frequent in SBP-patients (38.1%) than in cirrhotic controls (67.8%,P=0.021). CONCLUSION:In patients with alcoholic liver cirrhosis,the-2518 MCP-1 genotype AA is a risk factor for the development of SBP.

Fractalkine and TGF-β1 levels reflect the severity of chronic pancreatitis in humans

MM： To clarify whether serum chemokine and cytokine levels can become useful biological and functional markers to assess the severity of chronic pancreatitis （CP）. This study aimed at clarifying whether serum chemokine and cytokine levels can become useful biological and functional markers to assess the severity of CR METHODS： Serum monocyte chemoattractant protein-1 （MCP-1）, transforming growth factor beta-1 （TGF-βI）, and soluble type fractalkine （s-fractalkine） concentrations were examined in patients with CP （n = 109） and healthy controls （n = 116）. Severity of disease was classified in patients with CP by a staging system. Relationships between stage-specific various clinical factors and serum MCP-1, TGF-β1, and s-fractalkine levels were investigated. Furthermore, 57 patients with non-alcoholic CP were similarly evaluated in order to exclude influence of alcohol intake. RESULTS： Patients with CP showed significant higher levels of serum TGF-β1 and s-fractalkine, but not MCP-1, compared to the controls. Serum TGF-β1 in the severe stage and s-fractalkine in the mild and thesevere stage of CP significantly increased compared to those of controls. However, it was observed that both TGF-β1 and s-fractalkine levels were affected by alcohol intake. In patients with non-alcoholic CP, serum TGF-β1 showed significant increase in the moderate stage of CP, and serum s-fractalkine revealed significant increase in the early stage of CP. CONCLUSION： It is suggested that the measurement of serum F-fractalkine is useful to diagnose early- stage CP. Moreover, the combined determination of both, s-fractalkine and TGF-β1, in human sera may be helpful in evaluating the severity status of CP.

Predictive factors of clinical response in steroid-refractory ulcerative colitis treated with granulocyte-monocyte apheresis

AIM:To identify factors predicting the clinical response of ulcerative colitis patients to granulocyte-monocyte apheresis (GMA). METHODS:Sixty-nine ulcerative colitis patients (39 F,30 M) dependent upon/refractory to steroids were treated with GMA. Steroid dependency,clinical activity index (CAI),C reactive protein (CRP) level,erythrocyte sedimentation rate (ESR),values at baseline,use of immunosuppressant,duration of disease,and age and extent of disease were considered for statistical analysis as predictive factors of clinical response. Univariate and multivariate logistic regression models were used. RESULTS:In the univariate analysis,CAI (P = 0.039) and ESR (P = 0.017) levels at baseline were singled out as predictive of clinical remission. In the multivariate analysis steroid dependency [Odds ratio (OR) = 0.390,95% Confidence interval (CI):0.176-0.865,Wald 5.361,P = 0.0160] and low CAI levels at baseline (4 < CAI <7) (OR = 0.770,95% CI:0.425-1.394,Wald 3.747,P = 0.028) proved to be effective as factors predicting clinical response. CONCLUSION:GMA may be a valid therapeutic option for steroid-dependent ulcerative colitis patients with mild-moderate disease and its clinical efficacy seems to persist for 12 mo.

Lipid-associated membrane proteins of Mycoplasma penetrans induce production of proinflammatory cytokines in human monocytic cells

The aim of this study is to explore potential pathogenicity of Mycoplasma penetrans, and to investigate whether M. penetrans lipid-associated membrane proteins （LAMPs） could induce human monocytic cell line （THP- 1 ） to produce some proinflammatory cytokines in vitro, including interleukin- 1β （ IL- 1β）, tumor necrosis factor alpha （TNF-α）, and IL-8. THP-1 was stimulated with different concentrations of M.penetrans LAMPs and at different time to analyze the production of human IL-1β, TNF-α and IL-8. The protein levels of human IL-1β, TNF-α and IL-8 were measured by enzyme-linked immunoadsorbent assay （ELISA） and the mRNA levels of these proinflammatory cytokines were detected by reverse transcriptase-PCR （RT-PCR）. It was demonstrated in the present study that the production of IL-1β, TNF-α and IL-8 increased in dose- and time-dependent manner after stimulation with M. penetrans LAMPs in THP-1 cells. M. penetrans LAMPs also induced the expression of IL-1β, TNF-α and IL-8 mRNA. The production of IL-1β, TNF-α and IL-8 and the expression of mRNA were down-regulated by pyrrolidine dithiocarbamate （PDTC）. This study demonstrated that M. penetrans LAMPs can induce the production of proinflammatory cytokines in human monocytic cells in vitro, thus suggesting that it may be an important etiological factor.

Monocytes and Alzheimer's disease

Alzheimer＇s disease（AD） is a neurodegenerative disease characterized by extracellular amyloid beta（Aβ） deposition and intracellular neurofibrillary tangle formation.Monocyte is part of the innate immune system and can effectively remove dead cells and debris.It has been suggested that Aβ can recruit monocytes into brain in AD mice,resulting in restriction of cerebral amyloidosis.However,monocyte may act as a double-edged sword,either beneficial（e.g.,clearance of Aβ） or detrimental（e.g.,secretion of neurotoxic factors）.In addition,recent studies indicate that in AD patients,Aβ phagocytosis by monocytes is ineffective.The present review mainly summarized the current knowledge on monocytes and their potential roles in AD.

Effect of penehyclidine hydrochloride on patients with acute lung injury and its mechanisms

Objective： To assess the effects of penehyclidine hydrochloride on patients with acute lung injury （ALI）, to observe the expression of Toll-like receptor 4 （TLR4） on the peripheral monocytes of ALI patients and changes of inflammatory ＆ anti-inflammatory cytokines and to investigate the mechanism of TLR4 in ALI.Methods： Forty-five patients with ALI were randomly divided into penehyclidine hydrochloride treatment group （P group, n=21） and conventional treatment group （control group, C group, n=24）. Patients in both groups received conventional treatment, including active treatment of the primary disease, respiratory support, nutritional support and fluid management therapy, while those in P group were given penehyclidine hydrochloride （1 mg, im, q. 12 h） in addition.The TLR4 expression of 20 healthy volunteers were detected.The clinical effect, average length of stay in ICU and hospital,values of PaO2 and PaO2/FiO2, expression of TLR4 on the surface of peripheral blood mononuclear cells and some serum cytokines were evaluated for 48 h.Results： The general conditions of the two groups were improved gradually and PaO2 increased progressively.Compared with 0 h, PaO2 and PaO2/FiO2 at 6, 12, 24 and 48 h after treatment were significantly increased （P＜0.05）. The improvement in P group was obviously greater than that in C group （P＜0.05）. The average length of hospitalization showed no difference between the two groups, but penehyclidine hydrochloride significantly decreased the average length of stay in ICU （t=3.485, P＜0.01）. The expression of TLR4 in two groups were both obviously higher than that of healthy volunteers （P＜0.01）. It decreased significantly at 24 h （t=2.032, P＜0.05） and 48 h （t=3.620, P＜0.01）and was lower in P group than in C group. The patients who showed a higher level of TLR4 expression in early stage had a worse prognosis and most of them developed acute respiratory distress syndrome （ARDS）. The incidence of ARDS was 23.8% in P group and 29.17% in C group at 24 h.Until148 h, there were other two patients developing ARDS in control group. Serum IL-l, IL-8 and TNF-α expressions reduced after 24 h in both groups. The reduction in P group was more obvious than that in C group （P＜0.05）. IL-13 increased gradually from 0 h to 24 h, and decreased slightly at 48 h, which showed no difference between two groups （t=1.028, P＞0.05）.Conclusions： Penehyclidine hydrochloride improves the arterial oxygen pressure, down-regulates the expression of TLR4 and restrains the inflammatory cytokines in the downstream of TLR4 signaling pathway. It prevents the development of ALI and can be considered as an important drug in ALI treatment.

Dynamic activity of NF-κB in multiple trauma patients and protective effects of ulinastain

Objective： To investigate the dynamic activity of NF-κB at the early stage of injury in mul tiple trauma patients and the protective effects of ulinastain. Methods： From January 2008 to May 2010, patients with multiple traumas admitted to our emergency department were enrolled in this study. Their age varied from 20- 55 years. All enrolled patients were assigned randomly into control group （26 cases of multiple injury without ulinastain treatment）, ulinastain group （25 cases of multiple injury with ulinastain treatment）, and mild injury group （20 cases） for basic control. The inclusion criteria for mild injury group were AIS-2005 ≤ 3, single wound, previously healthy inhospital patients without the history of surgical intervention. In addition to routine treatment, patients in ulinastain group were intravenously injected 200 000 IU of ulinastain dissolved in 100 ml of normal saline within 12 hours after injury and subsequently injected at the interval of every 8 hours for 7 days. NF-κB activity in monocytes and the level of TNF- α, IL-1, IL-6 in serum on admission （day 0）, day 1, 2, 3, 4, and 7 were measured. Data were compared and analyzed between different groups. Results： NF-κB activity in monocytes and TNF- α, IL-1 and IL-6 of these patients reached peak levels at 24 hour after trauma, with gradual decrease to normal at 72 hour after trauma. NF-κB activity and levels of TNF- α, IL- 1 and IL-6 were lower in ulinastain group than control one, without any significant difference between the two groups. The mean duration for systemic inflammatory response syndrome and multiple organ dysfunction syndrome was 7 d±3.1 d and 10 d±3.5 d in ulinastain group and control group respectively, and showed a significant difference. Conclusions： NF-κB activity in monocytes and the levels of inflammatory cytokines in multiply injured patients increased transiently at the early stage of trauma. Ulinastain may shorten the duration of systemic inflammatory response syndrome and multiple organ dysfunction syndrome, but does not show the ability to decrease the activity of NF-κB.