人胰岛素原DNA疫苗预防非肥胖糖尿病鼠胰岛β细胞凋亡的机制探讨

目的探讨人胰岛素原DNA疫苗预防非肥胖糖尿病（NOD）小鼠胰岛β细胞凋亡的机制。方法（1）4周龄NOD雌鼠随机分为磷酸盐缓冲液（PBS）、质粒载体PcDNA3．1（PcDNA）、胰岛素原（Pins）3组，由胫前肌分别注射PBS、质粒PcDNA3．1、人胰岛素原DNA疫苗50pg，1周后重复1次。（2）各组取12周龄未发病的NOD鼠10只，分离胰腺，HE染色观察胰岛炎；TUNEL＋SABC法检测胰岛β细胞凋亡；取脾制成细胞悬液培养72h，ELISA法测定血清白介素4（IL-4）和γ干扰素（IFN-γ）水平；MTT法检测脾细胞增殖反应。结果（1）12周龄时Pins组正常胰岛比例高于PBS组和PcDNA组（均P〈0．01），胰岛周围炎比例低于PcDNA组（P=0．02），胰岛内炎比例则低于PBS组（P〈0．01）和PcDNA组（P=0．006）；（2）Pins组胰岛β细胞凋亡率低于PBS组和PcD—NA组（均P〈0．01）；（3）PIns组血清IL-4／IFN-γ比值显著高于PBS组（P=0．014）和PcDNA组（P=0．036）；（4）PIns组脾细胞对人胰岛素增殖反应的刺激指数（SI）低于PcDNA组（P=0．021）。结论人胰岛素原DNA疫苗通过上调IL-4／IFN-γ比值，使免疫平衡向Th2偏移，致NOD鼠胰岛炎减轻，胰岛β细胞凋亡减少。     

己酮可可碱预防NOD鼠1型糖尿病的机理研究

目的 探讨己酮可可碱（Pentoxifylline,PTX)对非肥胖糖尿病（NOD）小鼠1型糖尿病发病率，胰岛素的影响及其机制。方法 采用动物模型NOD鼠，注射环磷酰胺（CP）加速其发病。给PTX药物后计算糖尿病发病率，HE染色观察胰岛炎，并用逆转录（RT）PCR法检测脾细胞干扰素γ（IFN－γ），肿瘤坏死因子α（TNF－α），白介素10（IL－10）mRNA的表达。结果 PTX组糖尿病发生率（30．00％）明显低于对照组（67．86％）（P＜0．1）；胰岛炎程度也明显减轻（P＜0．001）；脾细胞IFN－γ，TNF－αmRNA的表达较对照组明显降低（P＜0．05），IL－10mRNA的表达无明显改变。结论 PTX可预防NOD鼠发生糖尿病，其机制可能与纠正Th1与Th2型细胞因子比例失衡有关。     

胰岛素预防NOD鼠糖尿病的机制研究

目的观察胰岛素（Ins）对非肥胖糖尿病（NOD）小鼠胰岛炎和糖尿病的影响，并探讨其诱导免疫耐受的机制。方法60只NOD雌鼠随机分为胰岛素（NPH）处理组和PBS对照组，分别于第4、12、20、28周予皮下注射NPH6U（60μl）及PBS60μl十不完全弗氏佐剂60μl。于12周龄观察胰岛炎和胰岛β细胞凋亡；测定血清和脾细胞上清IL-4和干扰素（IFN）γ浓度；检测胰岛内和脾细胞IL-4、IFN-γ mRNA的表达；并行联合过继转移实验。结果Ins组糖尿病发病率和β细胞凋亡率比PBS组低。血清和脾细胞上清IL-4浓度及胰岛内和脾细胞IL-4mRNA表达较PBS组高，而IFN-γ浓度及IFN—γ mRNA表达比PBS组低。过继转移实验Ins组DM发病率比PBS组低。结论Ins能诱导NOD鼠调节T细胞的产生，使全身和胰岛局部T细胞由Th1向Th2转型，抑制β细胞凋亡，从而预防DM．     

IL—4对NOD小鼠1型糖尿病免疫干预及其机制的探讨

目的：探讨IL－4对NOD鼠1型糖尿病发病率，胰岛炎的影响及其机制。方法：采用人类1型糖尿病动物模型NO（nonobese diabetic)鼠，IL－4处理后检测血糖，尿糖及糠愕病发病率，HE染色观察胰岛炎程度，流式细胞仪测定周围及中枢淋巴细胞亚群的变化，结果IL－4处理组糖尿病发病率为12．50％，明显低于对照组（62．50％）（P＜0．05），且胰岛炎的严重程度亦低于对照组；处理组周围单个核细胞（CD 4/CD 8及IL－2R＋细胞百分比增高，胸腺CD＋4，CD＋8细胞百分比增高而D＋4 CD＋8细胞百分比下降（P＜0．01）。结论 IL－4有降低NOD鼠1型糖尿病发病率及胰岛炎严重程度的作用，这种作用可能与IL－4影响了淋巴细胞亚群的分布从而纠正了免疫调节失衡有关。     

己酮可可碱对NOD小鼠1型糖尿病的影响

目的 探讨己酮可可碱（pentoxifyline,PTX)对NOD（non-obese diabetic)小鼠1型糖尿病的影响及其机制。方法 采用动物模型NOD鼠，PTX处理后检测血糖、尿糖及糖尿病发病率，HE染色观察胰岛炎，并用RT－PCR法检测胰腺干扰素γ（IFN-γ）、肿瘤坏死因子α（TNF－α）、白介素10（IL－10）mRNA的表达。结果 PTX组糖尿病发生率(30%)低于对照组（67．9％），P＜0．01；胰岛炎程度也减轻P＜0．001；胰腺IFN－γ、TNF－α mRNA的表达较对照组降低，P＜0．05；3项变化均具显著性。而IL－10 mRNA的表达则无显著改变。结论 PTX可预防NOD小鼠发生糖尿病，其机制可能与纠正Th1与Th2型细胞因子比例失衡有关。     

NOD鼠口服胰岛素诱导免疫耐受的疗效及形态学观察

目的 评价口服胰岛素诱导免疫耐受在防治自身免疫性糖尿病方面的效果。方法 雌性NOD鼠120只，随机分为胰岛素组和对照组各60只，6周龄始分别口服猪胰岛素500ul或磷酸盐缓冲液（PBS）500ul，至40周龄。6、12、20、25、30周龄分别测定两组未发病NOD鼠血清胰岛素（insulin)水平，并观察胰岛形态学变化。结果 40周龄糖尿病累计发生率对照组为91％，胰岛素组为11％，两者具有显著性差异（P＜0．01）。未发病NOD鼠中，20、25、30周龄对照组胰岛炎明显重于胰岛素组，血清胰岛素水平显著低于胰岛素组（P均＜0．05）。发病的NOD鼠中，对照组NOD糖尿病鼠血清胰岛素水平显著低于胰岛素组糖尿病鼠（P＜0．05）。结论 口服胰岛素可明显延迟NOD鼠糖尿病的发病时间，降低糖尿病发生率，缓解动物发病后的病情。     

吡格列酮预防非肥胖糖尿病鼠胰岛β细胞凋亡的机制研究

目的 探讨吡格列酮预防非肥胖糖尿病小鼠胰岛β细胞凋亡的机制.方法 (1)将4周龄NOD雌鼠分为吡格列酮(21只)及对照(21只)组,分别摄食含0.02％吡格列酮的混合饲料和普通营养饲料.观察52周龄的累积糖尿病发病率.(2)各组取12周龄未发病NOD鼠(n=15)胰腺,HE染色观察胰岛炎;TUNEL+SABC法检测胰岛β细胞凋亡.(3)ELISA法测定血清、脾细胞培养上清IFN-γ和IL-4水平及培养脾细胞核因子PPARγ、NF-κB活性.结果 (1)30、52周龄时,吡格列酮及对照组发病率分别为57.1％和76.2％ 、76.2％和90.5％(均P＞0.05);15周龄时,吡格列酮及对照组发病率分别为4.8％和33.3％(P=0.045).(2) 12周龄时,吡格列酮组正常胰岛和胰岛周围炎比例(14.73％,26.02％)高于对照组(5.69％,15.72％;均P＜0.01),胰岛内炎比例(59.25％)则低于对照组(78.59％,P＜0.01);吡格列酮组胰岛β细胞凋亡率(6.17％±3.62％)低于对照组( 10.62％±4.43％,P=0.008).(3)12周龄NOD鼠吡格列酮组血清IFN-γ水平[(561.05 ±78.61) pg/ml]显著低于对照组[(666.43±28.42) pg/ml,P=0.045];在培养的脾细胞上清中,吡格列酮组IFN-γ水平[(605.84±65.60) pg/ml]显著低于对照组[(692.20±44.98)pg/ml,P=0.041].(4)在培养的脾细胞中,吡格列酮组PPARγ活性(0.06±0.01)高于对照组(0.03±0.01,P=0.013),NF-κB活性(0.03±0.01)较对照显著降低(0.08±0.01,P=0.001).结论 吡格列酮活化PPARγ,抑制NF-κB活性,血清和脾细胞上清IFN-γ下降,Th细胞向Th1方向分化减少,NOD鼠胰岛炎减轻、胰岛β细胞凋亡减少.     

完全弗氏佐剂预防NOD鼠糖尿病的机制及其与细胞因子Th表达和β细胞凋亡的关系

目的　探讨完全弗氏佐剂 (CFA)预防非肥胖糖尿病 (NOD)小鼠胰岛炎与糖尿病的作用机制。　方法　 42只NOD雌鼠随机分为CFA处理组 (n =2 1)和磷酸盐缓冲液 (PBS)对照组 (n =2 1) ,3周龄时分别于后脚板一次性注射 50 μlCFA和PBS。于 12周龄时观察胰岛炎的程度、胰岛Fas和FasL的表达、β细胞凋亡情况以及测定血清白细胞介素 4(IL 4)、γ干扰素 (Ifn γ)水平和胰岛内IL 4、Ifn γ、白细胞介素 1β(IL 1β)、FasmRNA的表达水平。 　结果　 12周龄时CFA组胰岛炎积分和β细胞凋亡率均显著低于PBS组 (P均 <0 .0 5)。Fas只在PBS组胰岛内表达 ,而FasL在CFA和PBS组均有相似表达。 12周龄CFA组血清IL 4水平较PBS组增高而Ifn γ水平降低 (P均 <0 .0 5) ;RT PCR结果显示 ,CFA组胰岛内IL 4mRNA转录水平较对照组显著增高 (P <0 0 1) ,而Ifn γ、IL 1β、FasmRNA转录水平则明显降低 [P均 <0 .0 1(Ifn γ ,IL 1β) ,P <0 .0 5(Fas) ]。 3 0周龄时CFA组糖尿病发病率 (9.1% ,1/ 11)比PBS组 (81.8% ,9/ 11)显著降低 (P =0 .0 0 1)。　结论　CFA预防NOD鼠胰岛炎及糖尿病可能与胰岛自身反应T细胞由Th1向Th2转型 ,从而抑制Fas介导的 β细胞凋亡相关     

053 白介素-13预防NOD大鼠自身免疫性糖尿病

[英]/Zaccone P…//Diabetes.-1999,48.-1522～1528 研究了应用抗炎症细胞因子--重组人白细胞介素-13(hIL-13)治疗非肥胖糖尿病(NOD)大鼠自发性或转移性1型糖尿病发生的效果。 研究设计和方法本研究使用血糖正常的雌性NOD大鼠观察hIL-13对自发性1型糖尿病发生的影响，使用血糖正常的雄性NOD大鼠观察hIL-13对转移性1型糖尿病发生的影响。另有一组经磷酸盐缓冲溶液处理为对照组。分离大鼠脾脏的脾淋巴细胞(SLC)体外测定细胞因子的分泌；腹腔注射仓鼠抗-大鼠CD3单克隆抗体，体内测定细胞因子的分泌；应用酶联免疫吸附法(ELISA)测定肿瘤坏死因子-α(TMF-α)、干扰素-γ(IFN-γ)、白介素-4(IL-4)等细胞因子及免疫球蛋白IgE。胰岛组织学检查：根据胰岛单核细胞浸润程度分级评分，每只鼠至少计数15个胰岛，每个胰岛的平均分数为总分数除以所检查的胰岛数目，胰岛炎积分(ISs)用均数±标准差表示。 结果 本次实验应用hIL-13治疗白NOD鼠第5周龄开始至16周龄结束。实验结束时，hIL-13治疗组无一例发生1型糖尿病，而对照组Ⅰ型糖尿病的发生率为29％。组织学检查亦表明，治疗组比对照组胰岛炎较轻。急性糖尿病动物的脾细胞通过静脉注射转移到血糖正常的同源动物可引起该动物的自身免疫性糖尿病。本研究发现，脾细胞转移4周后，NOD大鼠对照组大部分发生了1型糖尿病，而hIL-13治疗组糖尿病发生率明显减少，组织学检查表明，治疗组ISs明显低于对照组(2.76±0.85比3.68±0.3，P=0.003)。鼠16周龄时处死并分离SLC，分析发现治疗组SLC比对照组分泌较少的IFN-γ。NOD鼠腹腔注射仓鼠抗大鼠CD3后治疗组鼠血TNF-α、IFN-γ、IL-4均低于对照组，且治疗组脾细胞转移的NOD大鼠血清IgE含量明显高于对照。 研究认为，hIL-13具有显著的免疫下调性质，从而进一步证明IL-4相关的抗炎性细胞因子可能在预防1型糖尿病的发生中起一定作用。 (杨秀英 李 萍摘 刘长山校)     

T细胞接种对环磷酰胺处理NOD小鼠糖尿病的影响

目的 探讨T细胞接种（TCV）对Ⅰ型糖尿病的预防作用。方法 将6周龄未发病的雌性NOD鼠用4周龄未发病、18周龄新近发病和32周龄发病时间长的NOD鼠脾细胞制得的T细胞疫苗接种,检测TCV对环磷酰胺处理的NOD鼠糖尿病发病率和胰岛炎程度的影响,以及接种后宿主淋巴细胞亚群的变化情况。结论 TCV可以降低糖尿病发病率、减轻胰岛单个核细胞的浸润程度,诱导宿主脾脏CD8^ T细胞百分比升高、CD4^＋／CD8^ 比值和IL－2R^ 细胞下降,胸腺CD4^- CD8^-单阳性细胞百分比升高。结论 TCV可降低宿主对自身免疫的反应性,这种作用可能与宿主脾脏、胸腺淋巴细胞亚群的变化和对糖尿病的预防效应有关。     

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.     

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.     

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.     

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.