脓毒症大鼠肠系膜淋巴结细胞免疫功能的变化

目的 探讨脓毒症大鼠肠系膜淋巴结细胞免疫功能的变化.方法 雄性Wistar大鼠96只,体重200～250 g,随机分为假手术组(S组)和盲肠结扎穿孔致脓毒症组(CLP组),每组48只.分别于CLP后即刻、6、12、24、48、72 h(T_(1～5))时随机取8只大鼠抽取腹主动脉血,采用鲎试剂偶氮显色法测定血浆内毒素水平;取肠系膜淋巴结组织,流式细胞仪测定肠系膜淋巴结组织Th1/Th2比值和调节性T淋巴细胞(Treg)占CD4~+细胞的比例(Treg比例).结果 与S组比较,CLP组T_(2～5)时血浆内毒素水平升高,T_1时Th1/Th2比值升高,Treg比例降低,T_(2～5)时Th1/Th2比值降低,Treg比例升高(P<0.05或0.01);Th1/Th2比值与Treg比例呈负相关(r=-0.87,P<0.05).结论 脓毒症大鼠肠系膜淋巴结细胞免疫功能降低,其机制可能与肠源性内毒素介导Treg比例增加有关.     

肝移植术后早期合并脓毒症患者的免疫功能障碍

目的 研究肝移植患者发生术后早期脓毒症时机体免疫状态的变化特点.方法 前瞻性选取2008年10月至2009年7月因慢性重型肝炎行肝移植手术患者19例,比较其与健康人群(20例)免疫状态的差异,并根据术后早期是否发生脓毒症分成脓毒症组(HSS组)和非脓毒症组(HSNS组),观察2组患者手术前后不同时期外周血T淋巴细胞亚群的动态变化.结果 与健康人群相比慢性重型肝炎患者T％、IFN-γ/IL-4均显著下降,CD4+ CD25+ Foxp3+ Treg(％)、Foxp3 mRNA 以及IL-10浓度明显上升.9例患者术后早期发生脓毒症,2组患者T％在术后第1天均较术前显著下降,HSNS组第14天恢复至术前水平,而HSS组仍维持在较低水平.HSNS组在术后第14天Treg％(t=3.265,P=0.004)和Foxp3mRNA表达(t=2.750,P=0.013)与术前相比明显降低,而HSS组术后则略呈上升趋势.HSNS组术后IFN-γ/IL-4有所上升,术后第3天最为显著(t=2.261,P =0.036),HSS组术后各时间点与术前比较无明显变化.两组患者IL-10浓度术后均明显下降,其中HSNS组一直维持在较低水平,而HSS组术后第14天再次上升至术前水平.结论 慢性重型肝炎患者免疫功能下降,术后未合并脓毒症者免疫状态于术后7～14d开始逐渐改善,发生脓毒症者仍维持术前的免疫低下状态.     

血必净促进内毒素/脂多糖刺激调节性T淋巴细胞凋亡并介导辅助性T淋巴细胞漂移的作用

目的 了解血必净注射液促进LPS刺激CD4+CD25+调节性T淋巴细胞(Treg细胞)凋亡过程及介导辅助性T淋巴细胞(Th)漂移的调节作用.方法 免疫磁珠法分选获得大鼠脾脏CD4+CD25+Treg细胞,分为常规培养对照组、抗CD3/CD28组、抗CD3/CD28+LPS组、抗CD3/CD28+血必净组和抗CD3/CD28+LPS+血必净组,培养3 d后应用流式细胞术检测Treg细胞凋亡率及叉头翼状螺旋转录因子3(Foxp3)表达.将CD4+CD25+Treg细胞与CD4+CD25+T淋巴细胞1:1培养,伴刀豆球蛋白A刺激68 h,检测上清液中Th1分泌的γ干扰素(IFN-γ)、Th2分泌的IL-4、Th17分泌的IL-17水平.结果 抗CD3/CD28+LPS+血必净组Treg细胞凋亡率为(45.1±2.7)%,明显高于抗CD3/CD28+LPS组[(29.4±1.6)%,P＜0.01];2组Foxp3平均荧光强度分别为95±9、140±18,差异有统计学意义(P＜0.01).同时,抗CD3/CD28+LPS+血必净组IFN-γ分泌水平显著高于抗CD3/CD28+LPS组(P＜0.01),IL-4则呈相反变化(P＜0.05),抗CD3/CD28+LPS+血必净组IFN-γ/IL-4较对照组升高(P＜0.01);抗CD3/CD28+血必净组IL-17分泌水平较抗CD3/CD28组明显下降(P＜0.05).结论 CD4+CD25+Treg细胞活化介导了Th1向Th2功能性极化;血必净对LPS诱导的T淋巴细胞免疫功能有重要调节作用,可促进CD4+CD25+Treg细胞凋亡并介导Th2向Th1漂移,从而缓解细胞免疫抑制状态.     

腹腔感染大鼠机体免疫状态变化

目的：观察腹腔感染大鼠的机体免疫状态变化,研究清热解毒方剂对机体细胞免疫失衡的影响。方法：采用人工胃液联合大肠杆菌腹腔内注射的方法建立大鼠腹腔感染模型,取Wistar大鼠120只随机分为对照组、模型组、清热解毒方剂组,造模后12h、24h、48h、72h经腹主动脉取血,采用流式细胞仪检测全血中辅助性T细胞1/2（Th1/Th2）比值以及调节性T细胞（Treg）比例的变化。结果：与正常对照组比较,模型组Th1/Th2比值在造模后12h明显升高,造模后48、72h明显降低（P〈0.05或P〈0.01）,而Treg比例在造模后12h明显下降,随后明显升高（P〈0.05或P〈0.01）,清热解毒方剂组Th1与Th2比值以及Treg比例升高和降低的幅度较模型组明显减低。在模型组和清热解毒方剂组,Th1/Th2比值与Treg比例呈线性负相关（P〈0.05）。结论：腹腔感染大鼠细胞免疫功能抑制,表现为Th1/Th2比值减少,其机制与肠源性内毒素介导Treg比例和活性增加有关;清热解毒方剂可能通过调节Treg比例来减少Th1向Th2的漂移,从而起到维持机体免疫平衡的作用。     

调节性T淋巴细胞与严重烧伤脓毒症

It has been demonstrated that severe burn per se may contribute to activation and proliferation of regulatory T cells (Treg). This characteristic phenomenon might allow Treg to function for a prolonged period of time to regulate immune response, and to induce suppression of T lymphocyte immune function. Different degrees of elevated levels of cytokines produced by Treg and activation markers on Treg surface could also be involved in the development of sepsis and fatal outcome in patients with severe burn. Thus, the regulation of Treg as a cellular therapeutic strategy might be important to the Th1/Th2 cytokine balance in burn patients complicated with sepsis.     

调节性T淋巴细胞与严重烧伤脓毒症

许多资料显示,大面积烧伤并发的脓毒症来势凶猛,病情进展迅速,患者病死率高,给临床救治工作带来极大困难.及时诊断、有效预防和治疗烧伤脓毒症,是提高严重烧伤救治成功率的关键.随着对感染病理生理过程认识的不断深化,人们对烧伤后脓毒症的发病机制有了新的了解.     

调节性T淋巴细胞与严重烧伤脓毒症

许多资料显示,大面积烧伤并发的脓毒症来势凶猛,病情进展迅速,患者病死率高,给临床救治工作带来极大困难.及时诊断、有效预防和治疗烧伤脓毒症,是提高严重烧伤救治成功率的关键.随着对感染病理生理过程认识的不断深化,人们对烧伤后脓毒症的发病机制有了新的了解.     

免疫调理对脓毒症大鼠中性粒细胞和淋巴细胞亚群的影响

目的研究免疫调理对脓毒症大鼠中性粒细胞和淋巴细胞及其亚群的影响,阐明免疫调理在脓毒症治疗中的作用。方法成功制作SD大鼠盲肠结扎穿孔（CLP）脓毒症模型,被分为假手术组、对照组和实验组,各20只。制作脓毒症模型后分别在3、12、24、48h采血待测。检查外周血涂片、白细胞总数、中性粒细胞、淋巴细胞计数和CD4^＋、CD8^＋T淋巴细胞亚群。结果实验组白细胞计数和中性粒细胞计数比对照组明显降低,而淋巴细胞计数和CD8^＋T淋巴细胞水平比对照组明显升高,P〈0．05。实验组CD8们林巴细胞水平与对照组差异无统计学意义。结论免疫调理可以明显降低脓毒症大鼠白细胞和中性粒细胞,从而减轻炎症反应。免疫调理可以提高脓毒症大鼠淋巴细胞计数和CD4^＋T淋巴细胞,从而改善免疫功能。     

动脉粥样硬化患者外周血CD4 T 细胞及CD4+CD25highCD127- 调节性T 细胞的变化

目的：探讨动脉粥样硬化（AS）患者外周血CD4 T细胞及调节性T细胞（CD4+CD25highCD127- Treg）比例及功能的变化。 方法：选择AS患者（AS组）及健康体检者（健康对照组）各40例,抽取外周静脉血,用流式细胞术测定CD4 T细胞、CD8 T细胞、IFN-γ+ CD4 T细胞、IFN-α+ CD4 T细胞和CD4+CD25highCD127- Treg细胞比例,以及各自的CD4+CD25highCD127- Treg细胞与CD4 T细胞共培养后,CD4 T细胞增殖能力以及CD4 T细胞分泌炎症因子能力的变化。 结果：与健康对照组比较,AS组外周血CD4 T细胞比例及CD4 T/CD8 T比值明显升高、IFN-γ+ CD4 T及IFN-α+ CD4 T细胞比例明显升高（均P<0.05）,而CD8 T细胞比例与CD4+CD25highCD127- Treg细胞比例差异无统计学意义（均P>0.05）。健康对照组CD4 T细胞的增殖能力、IFN-γ及IFN-α分泌能力在与CD4+CD25highCD127- Treg细胞共培养后较其单独培养均明显降低（均P<0.05）,而AS组CD4 T细胞的以上指标在与CD4+CD25highCD127- Treg细胞共培与单独培养间差异均无统计学意义（均P>0.05）。 结论：AS患者外周血CD4+CD25highCD127- Treg细胞功能的丧失,导致外周血CD4 T细胞比例升高、分泌炎症因子能力增强,从而引发AS的炎症状态。     

大面积烧伤脓毒症患者T淋巴细胞免疫功能的改变及临床意义

目的 了解大面积烧伤脓毒症患者T淋巴细胞免疫功能的变化，探讨其与脓毒症的关系。方法 选择59例烧伤面积≥30％TBSA的患者，分为脓毒症组43例和非脓毒症组16例。采集两组患者伤后1、3、5、7、14、21、28d的外周静脉血，检测T淋巴细胞增殖能力和白细胞介素2（IL-2）的分泌水平，并行相关性分析；通过流式细胞仪检测CD3^＋／CD4^＋ T淋巴细胞的百分率及其凋亡率，并行相关性分析。结果 与非脓毒症组比较，脓毒症组患者伤后l、14、2l、28dT淋巴细胞增殖能力和IL-2的分泌水平均显著下降（P〈0．05或P〈0.01），两指标呈显著正相关（r=0．82，P〈0．01）。伤后1、5、14、21、28d，脓毒症组患者CD3^＋／CD4^＋T淋巴细胞百分率明显低于非脓毒症组，而其凋亡率呈相反趋势（P〈0.05或P〈0．01），两指标呈显著负相关（r=-0.66，P〈0．05）。结论 大面积烧伤脓毒症患者T淋巴细胞免疫功能持续处于抑制状态，T淋巴细胞凋亡参与了脓毒症细胞免疫紊乱的病理生理过程。     

Prognostic Impact of Inferior Mesenteric Artery Lymph Node Metastasis in Colorectal Cancer

Aim

The aims of this study are to identify the natural course of inferior mesenteric artery (IMA) lymph node metastasis, and to evaluate the prognostic impact of IMA lymph node metastasis in the sigmoid colon and rectal cancer.

Patients and Methods

From our prospectively collected database, a total of 625 patients who underwent resection with curative intent for stage III adenocarcinoma of the sigmoid colon and rectal cancer between June 1995 and June 2007 were selected. Patients were divided into the IMA-positive group (n = 33) and the IMA-negative group (n = 592) according to IMA lymph node metastasis status. Clinicopathological features, recurrence patterns, and 5-year disease-free survival rates were compared between the two groups.

Results

Following curative resection, 5-year disease-free survival rate was 31.9% in the IMA-positive group and 69.4% in the IMA-negative group (p < 0.001). Cox regression analysis revealed that rectal cancer, pathologic stage, and presence of IMA lymph node metastasis were independently associated with disease-free survival. Systemic recurrence rate was significantly higher in the IMA-positive group than in the IMA-negative group (48.5 vs. 20.8%, respectively, p = 0.001). Para-aortic nodal recurrence showed significant association with presence of IMA lymph node metastasis on multivariate analysis (hazard ratio 11.8; 95% confidence interval 2.7–52.2, p = 0.001).

Conclusion

Presence of IMA lymph node metastasis should be considered as a predictive factor for high systemic recurrence, and should be treated and followed up with caution for para-aortic nodal recurrence.     

Surgical Management of Mesenteric Lymph Node Metastasis in Patients Undergoing Rectosigmoid Colectomy for Locally Advanced Ovarian Carcinoma

Background We sought to determine the incidence of mesenteric lymph node metastases in patients undergoing rectosigmoid resection for epithelial ovarian carcinoma and to evaluate the potential contribution of sigmoid mesocolectomy toward achieving complete surgical cytoreduction. Methods Pathology results for patients undergoing rectosigmoid colectomy for epithelial ovarian carcinoma from August 1998 through September 2005 were retrospectively reviewed. Fifty-three patients with pathological documentation of mesenteric lymph nodes were selected for further review. A focused analysis was performed on cases with an adequate surgical sampling of mesenteric lymph nodes (more than one positive or five total mesenteric lymph nodes) to determine the overall incidence of nodal metastases. χ² analysis was used to identify clinicopathologic factors associated with mesenteric lymphatic spread. Results A total of 39 (73.6%) of 53 patients had an adequate mesenteric resection suitable for nodal analysis. In this subgroup, 32 (82.1%) of 39 patients had one or more mesenteric lymph nodes containing metastatic ovarian carcinoma. Invasion beyond the serosa of the rectosigmoid colon was present in 31 (79.5%) of 39 of cases; however, increasing depth of invasion was not associated with risk of mesenteric nodal disease. In addition to bowel wall involvement, the only clinical factor that correlated with mesenteric lymph node involvement was concurrent tumor spread to retroperitoneal lymph nodes (P = .025). Conclusions Locally advanced ovarian carcinoma involving the rectosigmoid colon is associated with a high incidence of mesenteric nodal metastasis. Standard surgical technique should include a sigmoid mesocolectomy with resection of the associated lymphatic tributaries at the time of rectosigmoid colectomy if the surgical objective is complete cytoreduction of occult nodal disease.     

HELP Apheresis in the Treatment of Sepsis

Heparin induced extracorporeal lipoprotein fibrinogen precipitation (HELP) is an established procedure for removal of low-density lipoprotein (LDL) cholesterol, lipoprotein (a), and fibrinogen in patients with severe hypercholesterolemia. In vitro studies revealed that HELP also removes endotoxin, tumor necrosis factor α (TNF-α) and C-reactive protein (CRP). With the intention to treat, we applied this procedure to 4 patients with severe gram-negative sepsis with highly elevated endotoxin blood levels. Nine treatments were performed, 6 using the standard HELP precipitating buffer and 3 without addition of heparin to the precipitating buffer. Heparin was omitted from the precipitating buffer to avoid fibrinogen depletion in patients at risk (low fibrinogen, postoperative). The average processed plasma volume was 3,386 ml in the standard and 2,963 ml in the modified treatment. Mean reductions (%) in plasma solute concentrations were (standard/modified procedure) as follows: endotoxin, 50/57; TNF-α, 25/5; CRP, 49/55; fibrinogen, 49/6; total cholesterol, 38/5; and apolipoprotein B (Apo B), 41/2. Both treatment modalities were equally effective in removing endotoxin and CRP. With the modified precipitation buffer, fibrinogen was not removed. To further simplify the extracorporeal treatment, we have designed a closed-loop circuit with 2 adsorbers in series, one for removal of TNF-α (dextran sulfate modified cellulose) and the other for removal of endotoxin (DEAE-cellulose). In vitro evaluation confirmed very efficient endotoxin and TNF-α removal from plasma. This system is very simple, operates at physiological pH, and uses adsorbers already in clinical use for other purposes.     

Prognostic Value of Lymph Node Yield and Metastatic Lymph Node Ratio in Medullary Thyroid Carcinoma

Introduction  Lymphadenectomy and thyroidectomy is standard treatment for medullary thyroid carcinoma (MTC), but the prognostic importance of the number of lymph nodes removed (lymph node yield, LNY) and the proportion of metastatic lymph nodes resected (metastatic lymph node ratio, MLNR) is unknown. We hypothesized that MTC survival is influenced by LNY and MLNR. Methods  Patients (N = 534) who underwent thyroidectomy with lymphadenectomy for MTC between 1988 and 2004 were identified in the Surveillance, Epidemiology, and End Results (SEER) database. The Kaplan–Meier method was used for univariate comparisons of survival for LNY and MLNR with a maximum follow-up of 12 years. Cox regression models adjusted for age, sex, extent of disease, tumor size, nodal status, LNY, and MLNR. Results  By univariate analysis, increasing LNY was associated with improved survival in all patients (P < 0.002) and node-positive patients (P < 0.001). In a multivariate analysis using LNY and MLNR as categorical variables, significant factors influencing survival included: age (P < 0.001), tumor size (P < 0.001), LNY (P = 0.007), and MLNR (P < 0.02); in node-negative patients: age (P = 0.002); in node-positive patients: age (P < 0.001), tumor size (P < 0.001), and LNY (P = 0.001). Using LNY and MLNR as continuous variables, significant factors influencing survival included: age (P < 0.001), tumor size (P < 0.001), and MLNR (P = 0.01); in node-negative patients: age (P < 0.001); in node-positive patients: age (P < 0.001) and tumor size (P < 0.001). Conclusion  In patients undergoing thyroidectomy and lymphadenectomy for MTC, LNY and MLNR predict poorer survival, but their impact on survival was limited to node-positive patients and was otherwise dominated by the effects of age and extent of disease. Supported by Grant Number KL2RR024144 from the National Center for Research Resources. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Center for Research Resources or the National Institutes of Health. Presented in part at the 3rd Annual Academic Surgical Congress, February 12–15, 2008, Huntington Beach, California.