乳腺癌HER2／neu手性多肽疫苗的实验研究

目的研究具有局部手性的HER-2／neu多肽疫苗对细胞毒性T细胞（CTLs）的诱导作用。方法用固相法合成HER-2／neu手性肽疫苗并进行纯化，建立小鼠的乳腺癌模型，用上述手性肽做疫苗、重组人粒细胞．巨噬细胞集落刺激因子（GM-CSF）为佐剂，对小鼠进行免疫，分别应用MTT法、ELISPOT法、颗粒酶释放法测定T细胞的增殖，T细胞分泌IFN-γ，CTL对肿瘤细胞的杀伤活性。结果合成的手性HER-2／neu可明显的促进T细胞增殖，促进T细胞分泌IFN-γ及诱导CTL杀伤乳腺癌细胞。结论手性HER-2／neu多肽疫苗可在小鼠体内诱导特异细胞免疫，增加对乳腺癌细胞的杀伤作用。     

P356-1Y2L表位刺激BALB/c小鼠的特异性抗肿瘤免疫

目的检测MAGEC2衍生的免疫原性肽在小鼠体内抗肿瘤免疫效果。方法测定P356-1Y2L免疫BALB/c小鼠后诱导CTL以及分泌IFN-γ的能力,用MAGEC2衍生的表位免疫荷瘤小鼠,检测其抑制肿瘤生长的能力及观察小鼠的生存能力。结果 P356-1Y2L免疫BALB/c小鼠引发CD8~+T淋巴细胞的强烈增殖和活化,且能分泌较多的IFN-γ,并且可以有效的杀伤靶细胞。用P356-1Y2L免疫荷瘤小鼠,能够抑制小鼠肿瘤生长并延长小鼠的生存期。结论免疫原性肽P356-1Y2L能够在BALB/c小鼠中引发特异性CTL反应,成为癌症治疗性多肽疫苗的候选表位。     

白细胞介素-15重组体对HBsAg核酸疫苗的免疫佐剂作用

白细胞介素-15(IL-15)属螺旋结构细胞因子家族，它能促进T细胞、自然杀伤(NK)细胞和B细胞增殖及分化；具有诱导淋巴细胞产生γ干扰素(IFN-γ)及肿瘤坏死因子α的作用。IL-15与疫苗共同免疫时可刺激抗体产物增加，细胞毒性T淋巴细胞(CTL)活性加强。将IL-15表达质粒与HBsAg DNA疫苗共同免疫小鼠，以观察其对小鼠免疫应答的影响。     

共表达PRRSV ORF5和ORF6基因重组核酸疫苗对断奶仔猪的免疫效果研究

目的评价猪繁殖与呼吸综合征病毒(PRRSV)重组核酸疫苗pVAX-ORF5-2A-ORF6对断奶仔猪的免疫效果。方法分别以pVAX-ORF5-2A-ORF6+Quil A、pVAX-ORF5-2A-ORF6、pVAX空质粒和PRRSV弱毒疫苗对断奶仔猪进行免疫接种,检测各免疫组血清中特异性抗体效价、细胞因子IL-2、IL-4、IL-6、IFN-γ的水平、T淋巴细胞增殖程度和CTL杀伤活性,评价其免疫效果。结果重组核酸疫苗pVAX-ORF5-2A-ORF6能够刺激猪体产生PRRSV特异性抗体,促进致敏T淋巴细胞增殖,诱导产生特异性细胞毒性T淋巴细胞(CTL)杀伤活性;免疫仔猪血清IL-2和IFN-γ水平与对照组比较差异有统计学意义(P<0.05),加入免疫佐剂Quil A的实验组IFN-γ水平显著高于未加佐剂的实验组(P<0.05)。结论重组核酸疫苗pVAX-ORF5-2A-ORF6可诱导免疫仔猪产生特异性的体液免疫和细胞免疫应答,有望成为抗PRRSV感染的新型候选疫苗。     

重组IL-12逆转录病毒转染树突细胞体外对鼻咽癌细胞的杀伤作用

目的体外实验评估重组白细胞介素(IL)-12逆转录病毒转染树突细胞对鼻咽癌细胞的杀伤作用。方法构建重组IL-12重组逆转录病毒,体外转染树突细胞(DC),经培养后收集上清液,检测IL-12、干扰素-γ(IFN)的分泌水平。以鼻咽癌细胞CNE-2为靶细胞,进行细胞毒T淋巴细胞(CTL)细胞毒活性测定。结果成功构建含有m IL-12的重组逆转录病毒;IL-12基因修饰DC可以显著诱导的大量IL-12和IFN-γ分泌,与未转染DC组比较,差异有显著意义(P0.01);DC-IL-12诱导的CTL及其上清液对鼻咽癌细胞CNE-2均有显著杀伤作用,与未转染DC组相比较差别有显著(P0.01)。结论 IL-12基因修饰树突细胞可显著诱导大量IFN-γ分泌并增强其诱导特异性CTL对肿瘤的杀伤效能。     

结核分枝杆菌MPT64抗原DNA疫苗在小鼠体内诱导的免疫应答

目的研究结核分枝杆菌MPT64抗原DNA疫苗在小鼠体内诱导的免疫应答。方法用表达MPT64的真核表达质粒pcDNA-M免疫BALB/c小鼠,ELISA法检测免疫小鼠的特异性抗体滴度和抗体亚类。分离免疫小鼠的脾淋巴细胞,检测淋巴细胞增殖、IFN-γ和IL-12产生水平、流式细胞仪计CD4+细胞和CD8+细胞数、脾淋巴细胞特异性CTL杀伤效应。结果MPT64基因免疫可诱导小鼠高水平的体液免疫应答,免疫小鼠脾淋巴增殖显著,IFN-γ和IL-12含量增加,CD4+细胞和CD8+细胞百分比明显增加,CTL杀伤效应明显。结论MPT64 DNA疫苗可诱导小鼠有效的体液和细胞免疫应答,有可能作为新型TB疫苗的组分。     

负载MB49肿瘤细胞抗原的DC疫苗对膀胱癌的抑制效应及机制

目的探讨负载MB49肿瘤细胞抗原的树突状细胞疫苗（DC疫苗）治疗膀胱癌的可行性及机制。方法采用反复冻融法提取MB49抗原并用其致敏DC2.4,制备DC疫苗。将24只膀胱癌荷瘤小鼠随机分为两组各12只。DC组皮下注射DC疫苗,对照组注射PBS。注射3周观察两组肿瘤体积变化,计算肿瘤抑制率;分离两组脾脏T淋巴细胞进行培养,MTT法检测细胞毒性T淋巴细胞（CTL）对MB49的杀伤率;ELISA法检测培养上清中IFN-γ水平。结果 DC组肿瘤抑制率及细胞杀伤率均明显高于对照组,P〈0.01;IFN-γ水平亦明显高于对照组,P〈0.01。结论以DC2.4为基础的DC疫苗体内具有抑制肿瘤细胞MB49增长的作用;其机制可能为激活T淋巴细胞。     

恶性疟原虫FCC1／HN株CSP抗原细胞免疫应答IL-2和IFN-γ的体外诱生分析

本实验利用恶性疟原虫FCCI/HN株CSP抗原基因在真核细胞(HeLa)中高效表达,将纯化的表达蛋白作为候选疫苗免疫小鼠,并通过体外诱生检测小鼠脾细胞白介素2(IL-2)和γ干扰素(IFN-γ)水平。结果　显示经疫苗免疫后的BALB/C小鼠脾细胞体外经特异性刺激后产生的IL-2和IFN-γ水平有显著提高。与我们前期研究结果　比较,表明对该疫苗特异性的IL-2、IFN-γ的产生与淋巴细胞增殖以及NK细胞活性增高有明显的相关性;该疫苗对T细胞有一定的刺激能力,可诱导小鼠产生较强的细胞免疫反应。     

DNA疫苗诱导小鼠特异性细胞免疫及抗乙型肝炎病毒皮下移植瘤的研究

目的观察乙型肝炎病毒(HBV)DNA疫苗(pCR3 1-S)诱导Balb/c小鼠(H-2d)的特异性细胞免疫应答及其对稳定表达HBsAg的小鼠肥大细胞瘤P815细胞(P815 HBV-S)(H-2d)成瘤性的影响.方法肌肉注射DNA疫苗,背部皮下接种P815-HBV S细胞,观察成瘤情况,4h 51Cr释放法检测小鼠脾细胞细胞毒T细胞(CTL)杀伤活性.结果DNA疫苗可以降低成瘤率,抑制肿瘤生长,延长小鼠平均存活期,提高小鼠存活率.CTL细胞杀伤活性明显增加(P＜0.001).结论DNA疫苗可以诱导细胞免疫应答,对体内HBV感染可能具有预防及治疗作用.     

尿酸辅助树突细胞免疫对小鼠淋巴细胞增殖及CTL效应的影响

目的：观察尿酸辅助HBsAg蛋白负载的树突细胞免疫接种对小鼠免疫功能的影响。方法：将负载HB-sAg的小鼠骨髓来源树突细胞经尾静脉注射接种小鼠，1次／w，共2次。分DC（树突状细胞）对照组、联合尿酸组、尿酸对照组。MTT法检测体外经HBsAg或PBS重刺激的脾T淋巴细胞增殖反应；流式细胞仪法检测CTL（细胞毒性T淋巴细胞）活性。结果：免疫2周时，小鼠脾T细胞增殖反应及体内HBsAg特异性CTL的杀伤活性，联合尿酸组明显强于各对照组。结论：尿酸可促进负载HBsAg树突细胞免疫后小鼠脾T淋巴细胞的增殖及HBsAg特异性CTL效应。尿酸具有增强DC疫苗免疫效应的活性，可用作研制抗HBV的治疗性疫苗的免疫佐剂。     

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.