慢性持续期支气管哮喘患者外周血中性粒细胞和单核细胞Toll样受体的表达及意义

目的探讨慢性持续期支气管哮喘（简称哮喘）患者外周血中性粒细胞和CD14＋成熟单核细胞TLR2、TLR4表达的变化及意义。方法入选慢性持续期哮喘患者30例,健康对照组46名,应用流式细胞仪全血双色免疫荧光直标法检测中性粒细胞和单核细胞TLR2和TLR4的表达情况,结果以细胞的TLR相对平均荧光强度（rmfi）和TLR相对阳性细胞百分率（rpcp）表示。结果哮喘组患者中性粒细胞TLR2表达的rmfi（1.48±0.19）和rpcp（M=1.34%,Q1-Q3为0.65%～2.91%）均明显低于健康对照组[分别为1.99±0.53和14.13%（Q1-Q3为2.61%～27.51%）,P值均〈0.001];TLR4表达的rmfi（1.77±0.21）明显低于对照组（1.95±0.26）,（P〈0.01）,rpcp（8.16±5.03）%与对照组（9.58±6.40）%比较,差异无统计学意义（P〉0.05）。哮喘组CD14＋单核细胞中TLR2 rpcp（90.92±8.42）%和TLR4 rpcp（11.92±10.34）%明显高于健康对照组[分别为（78.52±18.79）%和（5.78±3.98）%,P值均〈0.01];TLR2 rmfi（9.91±2.14）及TLR4 rmfi（2.60±0.74）与对照组[分别为（9.73±2.71）和（2.77±0.58）]差异无统计学意义（P值均〉0.05）。结论慢性持续期哮喘患者外周血中性粒细胞TLR2和TLR4的表达较健康人群明显下调,CD14＋单核细胞TLR2及TLR4阳性细胞率显著高于健康人群,中性粒细胞和单核细胞TLRs表达的变化在哮喘病程中可能发挥不同的免疫作用。     

原发性胆汁性肝硬化患者外周血单个核细胞Toll样受体表达与调节性T细胞相关性分析

目的 探讨原发性胆汁性肝硬化(PBC)患者外周血单个核细胞(PBMC)Toll样受体2、4、9(TLR2,TLR4,TLR9)表达及PBMC中CD4+ CD25+调节性T细胞(Tregs)特点和影响因素.方法 采用流式细胞术检测PBC患者(52例)和健康对照者(22例)外周血PBMC中TLR2、4、9阳性细胞及Tregs比例,比较其在PBC患者和健康对照者中的差别并分析TLR与Tregs相关性.结果 PBC患者Tregs比例低于健康对照者[(1.53+1.33)vs(4.42±1.43),P＜0.0001],TLR4阳性细胞比例高于健康对照者[(36.95±3.53)vs(32.84±8.06),P=0.003];PBC患者Tregs比例与TLR9阳性细胞比例呈负相关(R2 =0.115,P=0.016),健康对照者Tregs比例与TLR2,4和TLR2,4,9联合表达阳性细胞比例呈负相关(R2=0.326,P=0.007;R2 =0.226,P=0.034).结论 PBC患者外周血PBMC中Tregs比例降低,TLR4表达升高,可能受肝硬化失代偿期腹腔感染的影响.     

高血压病患者外周血单核细胞Toll样受体4的表达

目的 探讨Toll样受体4(TLR4)在高血压伴靶器官损害患者外周血白细胞上的表达及其表达差异在高血压发病中的意义.方法 采用流式细胞仪检测45例轻型高血压不伴靶器官损害患者、40例高血压伴靶器官损害患者及40名健康对照者外周血淋巴细胞、单核细胞和中性粒细胞上TLR4的表达.结果 3组外周血淋巴细胞和中性粒细胞TLR4的阳性率平均值低于6%;高血压不伴靶器官损害组单核细胞的TLR4阳性率为(48.2±14.3)%,高血压伴靶器官损害组(55.9±15.2)%,均显著高于健康对照组的阳性率[(40.8±13.6)%,P均＜0.01),高血压伴靶器官损害组的TLR4阳性率高于高血压不伴靶器官损害组(P＜0.05).结论 TLR4主要表达于外周血单核细胞,在高血压及伴靶器官损害患者其阳性率升高;TLR4及其介导的炎性反应与高血压的发生发展存在一定的相关性.     

2型糖尿病患者单核细胞TLR4表达及其与血浆TNF-α相关性研究

目的 探讨2型糖尿病患者单核细胞TLR4表达的变化及其与血浆炎症因子TNF-α的关系.方法 应用实时定量PCR及流式细胞仪检测22例2型糖尿病患者及20例健康对照者外周血单核细胞TLR4mRNA表达及TLR4阳性单核细胞比率,应用ELISA法检测血浆TNF-α浓度.结果 2型糖尿病患者外周血单核细胞TLR4mRNA相对表达量及TLR4阳性单核细胞比率为3.05±0.56、(14.93±3.48)%明显高于对照组1.48±0.30、(7.79±1.63)%(P＜0.05),血浆TNF-α浓度为(49.29±7.09)ng/L,高于对照组(39.55±5.22)ng/L(P＜0.05),TLR4阳性单核细胞比率与血浆TNF-α浓度正相关(r=0.543,P＜0.05).结论 2型糖尿病患者单核细胞TLR4表达增高,单核细胞TLR4可能参与了2型糖尿病的炎症反应.     

哈萨克族冠心病患者外周血单核细胞TLR4的水平及临床意义

目的探讨新疆哈萨克族(哈族)冠心病患者外周血单核细胞Toll样受体4(Toll-like Receptor4,TLR4)表达及与临床意义。方法根据冠状动脉造影结果及患者既往心脏病史,选取67例哈族冠心病患者为观察组,其中急性冠脉综合征患者33例,慢性冠脉病患者34例,选取经冠状动脉造影证实无冠心病者35例为对照组,采用流式细胞仪检测外周血淋巴细胞、单核细胞和中性粒细胞上TLR4的表达。结果 (1)各组外周血淋巴细胞和中性粒细胞TLR4阳性率平均值低于5%,单核细胞TLR4阳性率冠心病组显著高于对照组,差异有统计学意义(P0.01);(2)急性冠脉综合征组单核细胞TLR4阳性率高于慢性冠脉病组,差异有统计学意义(P0.05)。结论 TLR4主要表达于外周血单核细胞,TLR4阳性率与冠心病存在相关性,而且冠心病不同类型TLR4阳性率存在差异。     

冠心病患者PTCA前后外周血CD11b、CD62P表达变化及意义

采用流式细胞仪检测23例健康体检者（对照组）及41例冠心病患者（观察组）,经皮冠状动脉腔内成形术（PTCA）术前72、2h、术后30min及24、72h外周血中性粒细胞和单核细胞表面CD11b、CD62P的表达。结果观察组患者外周血中性粒细胞和单核细胞CD11b平均荧光强度（MFI）和CD62P阳性细胞百分比较对照组明显升高。PTCA术后24hCD11b荧光强度和CD62P阳性细胞百分比较术前72、2h明显升高。血小板CD62P与中性粒细胞CD11b呈正相关（r=0．253）。认为PTCA术后中性粒细胞和单核细胞CD11b及血小板CD62P表达上调,且可作为反映PTCA后炎症反应和血栓形成状况的指标。     

隐球菌性脑膜炎血和脑脊液白细胞人纤维介素的表达

目的 探讨人纤维介素(hFgl2)的表达与隐球菌性脑膜炎(CM)的关系.方法 采用免疫组织化学法检测20例CM患者(CM组)和20名健康者(健康对照组)血WBC及11例配对CM患者脑脊液WBC hFgl2的表达情况,并用医学影像分析系统(MIAS)对CM患者血和脑脊液hFgl2的表达进行灰度分析(图像的平均灰度值与hFgl2蛋白表达呈负相关).结果 CM组和健康对照组血单核细胞中hFgl2蛋白表达的平均灰度值分别为114.49±11.82、137.56±10.64,中性粒细胞中hFgl2蛋白表达的平均灰度值分别为111.71±8.62、143.56±12.57；与健康对照者比较,CM患者血单核细胞和中性粒细胞hFgl2的表达均明显升高(t值分别为21.21和27.19,均P＜0.01).CM患者和阴性对照脑脊液单核细胞中hFgl2蛋白表达的平均灰度值分别为113.33±8.07、147.25±9.72,中性粒细胞中hFgl2蛋白表达的平均灰度值分别为115.44±8.67、149.25±8.57；与阴性对照比较,CM患者脑脊液单核细胞和中性粒细胞hFgl2的表达均明显升高(t值分别为29.95和28.87,均P＜0.01).CM患者的血与配对脑脊液的单核细胞和中性粒细胞中hFgl2蛋白表达的差异均无统计学意义(t值分别为0.924和0.607,均P＞0.05).结论 hFgl2可表达于CM患者的血和脑脊液单核细胞和中性粒细胞.     

高血压病患者外周血单核细胞Toll样受体4的表达

目的探讨Toll样受体4(TLR4)在高血压伴靶器官损害患者外周血白细胞上的表达及其表达差异在高血压发病中的意义。方法采用流式细胞仪检测45例轻型高血压不伴靶器官损害患者、40例高血压伴靶器官损害患者及40名健康对照者外周血淋巴细胞、单核细胞和中性粒细胞上TLR4的表达。结果3组外周血淋巴细胞和中性粒细胞TLR4的阳性率平均值低于6%;高血压不伴靶器官损害组单核细胞的TLR4阳性率为(48.2±14.3)%,高血压伴靶器官损害组(55.9±15.2)%,均显著高于健康对照组的阳性率[(40.8±13.6)%,P均<0.01),高血压伴靶器官损害组的TLR4阳性率高于高血压不伴靶器官损害组(P<0.05)。结论TLR4主要表达于外周血单核细胞,在高血压及伴靶器官损害患者其阳性率升高;TLR4及其介导的炎性反应与高血压的发生发展存在一定的相关性。     

Toll 样受体在新生儿败血症患者中的动态变化及临床意义

目的：观察Toll样受体（TLR2、TLR4）在新生儿败血症病程中的动态变化,探讨其在新生儿抗感染免疫中的作用。方法将60例不同胎龄（31～42周）新生儿分为败血症组和对照组。应用流式细胞仪检测败血症组感染后第1天、第7～10天和非感染组第1天的外周血单核细胞和粒细胞表面TLR2、TLR4蛋白表达;同时检测血CRP、WBC、血小板。结果感染后第1天,败血症组TLR2蛋白在粒细胞和单核细胞的表达水平及CRP均高于非感染组（ P均＜0．05）;第7～10天,TLR2在粒细胞的表达水平接近对照组,而在单核细胞中则持续高表达（ P＜0．05）。败血症组TLR4在粒细胞表面持续低表达,在单核细胞表达高于粒细胞（ P＜0．05）;第7～10天,TLR4在单核细胞表达水平明显下降,接近对照组,但与第1天比P＜0．05。两组治疗前后WBC、血小板均变化不明显。结论新生儿败血症患儿外周血单核细胞和中性粒细胞表面TLR2、TLR4蛋白表达异常增高,提示TLR信号传导途径参与了新生儿的抗感染免疫机制,与新生儿败血症的发生、发展密切相关。     

高迁移率族蛋白1及Toll样受体4在类风湿关节炎患者血清和外周血单个核细胞中的表达及意义

目的 通过研究高迁移率族蛋白(HMGB)1及Toll样受体(TLR)4在类风湿关节炎(RA)外周血单个核细胞(PBMC)和血清中的表达,探讨HMGB1的作用及其与疾病活动的关系.方法 选取活动期RA患者38例和非活动期RA患者36例及健康对照组26名.采用反转录-聚合酶链反应(RT-PCR)方法检测PBMC中HMGB1 mRNA的表达,酶联免疫吸附试验(ELISA)检测血清中HMGBI蛋白的表达.采用流式细胞术分析PBMC上TLR4的表达.结果 ①活动期RA组PBMC中HMGB1 mRNA相对表达量和血清中HMGB1蛋白水平均高于健康对照组和非活动期RA患者[分别为2.63与0.71.0.93和(10.2±1.2)与(7.5+1.8),(8.3±1.8)ng/ml,P＜0.01.②活动期RA患者CD14+单核细胞和CD3+淋巴细胞上TLR4蛋白相对表达量高于非活动期和健康对照组(P＜0.05或P＜0.01),非活动期患者高于健康对照组(P＜0.05或P＜0.01).③血清中HMGB1蛋白水平与红细胞沉降率(ESR)、C反应蛋白(CRP)、类风湿因子(RF)、关节压痛数、肿胀数及关节X线分期均呈正相关,亦与TLR4蛋白相对表达量呈正相关.结论 RA患者PBMC具有合成和分泌HMGB1蛋白的功能,并部分通过与TLR4结合激活炎症反应,加重骨质破坏.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.