急性肝功能衰竭大鼠模型中程序性死亡-1及其配体的表达

目的 探讨程序性死亡-1(PD-1)/程序性死亡配体-1(PD-L1)在急性肝功能衰竭大鼠模型中的表达变化及其在肝脏急性炎性损伤中的作用.方法 SD大鼠分为2组:正常组6只,模型组30只,以D-氨基半乳糖(D-Gal)诱导急性肝功能衰竭大鼠模型.造模后分别在12、24、48、72和120 h取大鼠血及肝脏标本,采用RT-PCR法检测肝组织中PD-1 mRNA、PD-L1 mRNA的表达.计量资料组间比较用t检验,相关性检验用Pearson直线相关分析.结果 造模后12 h,大鼠血ALT、AST明显升高,分别为(217.3±33.7)U/L和(397.2±101.3)U/L,显著高于正常组的(30.5±3.1)U/L和(78.6±4.2)U/L,差异有统计学意义(t=-8.921,-6.121,均P＜0.01),至48 h达高峰.造模后12 h,模型组大鼠PD-1 mRNA表达(0.385±0.074)高于正常组(0.097±0.009),差异有统计学意义(t=-7.725,P＜0.01),48 h达高峰(0.927±0.132),72 h则明显下降.PD-L1mRNA在正常大鼠肝组织中表达很少,模型组PD-L1 mRNA水平逐渐升高,48 h达高峰(0.593±0.105)(t=-10.076,P＜0.01).造模后大鼠PD-1、PD-L1表达水平与血清ALT水平呈正相关(r=0.807,0.792,P＜0.01).结论 PD-1/PD-L1表达在急性肝功能衰竭大鼠肝脏炎性损伤中可能起重要作用.     

血清PCT水平、APACHEⅡ评分评估脓毒症患儿预后的临床价值

目的 探讨血清降钙素原(PCT)水平、急性生理学及慢性健康状况评分系统Ⅱ评分(APACHEⅡ评分)与脓毒症患儿预后的关系.方法 脓毒症患儿66例,均入儿童ICU(PICU)治疗.根据入PICU 7 d时患儿是否死亡分为存活组与死亡组.分析两组治疗前行APACHEⅡ评分;治疗前(t0)及治疗后1 d(t1)、4 d(t2)、7 d(t3)血清PCT水平.分析二者判断患儿预后的价值.结果 随着治疗的进程两组PCT均逐渐下降.从治疗第4天开始,PC与t0相比有明显下降(P＜0.01).至病程第7天,存活组PCT已接近正常水平;死亡组虽有下降趋势,仍持续高于正常水平.治疗前APACHEⅡ评分死亡组显著高于存活组(P＜0.01).PCT与APACHEⅡ评分的相关系数为0.721,P＜0.01.ROC曲线下面积值PCT水平为0.887,APACHEⅡ评分为0.888.二者联合评估,其敏感度高于单一指标.结论 血清PCT水平、APACHEⅡ评分与脓毒症患儿预后有较强的相关性;对二者进行综合分析可提高对脓毒血症预后评估的准确度.     

探讨GAS6在儿童下呼吸道感染性疾病中的诊断价值

目的:探讨GAS6在儿童下呼吸道感染性疾病和脓毒症诊断中的应用价值。方法:分别检测240例疾病组和55例正常对照组血清GAS6、PCT和CRP水平。比较各疾病组和正常对照组之间GAS6、PCT、CRP的差异。同时,绘制各诊断指标的ROC曲线。结果:脓毒症组、细菌性肺炎组和病毒性肺炎组患儿的血清GAS6均显著高于正常对照组(P0.05),但支原体肺炎组患儿的GAS6与正常对照组比较差异无统计学意义(P0.05)。脓毒症组患儿血清GAS6显著高于另外3组(P0.01),此外,细菌性肺炎组患儿血清GAS6明显高于支原体肺炎组和病毒性肺炎组(P0.05),但支原体肺炎组和病毒性肺炎组患儿间并无统计学差异(P0.05)。无论诊断脓毒症还是细菌性肺炎,GAS6的ROC曲线下面积均小于PCT和CRP(P0.05),但诊断病毒性肺炎时,GAS6与PCT的ROC曲线下面积差异无统计学意义(P0.05)。结论:在儿童下呼吸道感染性疾病中,患儿血清GAS6、PCT和CRP会显著升高。但GAS6对儿童下呼吸道感染性疾病及脓毒症的诊断价值不及PCT和CRP。     

地塞米松治疗与细菌性脑膜炎脑源性神经营养因子及其受体基因表达的关系

目的 探讨地塞米松(DEX)治疗与细菌性脑膜炎脑源性神经营养因子(BDNF)及其受体TrkB基因表达的关系。方法 建立细菌性脑膜炎模型，分别用抗菌药物及抗菌药物加DEX治疗，用原位杂交检测脑组织BDNF mRNA和TrkB mRNA的表达。结果 单独使用抗菌药物治疗后BDNF mRNA和TrkB mRNA表达均低于感染后5d组的水平(P＜0．05)，并以BDNF mRNA下降更明显(P＜0．01)，而使用DEX与抗菌药物联合治疗后，BDNF mRNA和TrkB mRNA表达回到较高水平(P＜0．01)，BDNFmRNA达到感染后5d组水平(P＞0．05)，TrkB mRNA则超过感染后5d组水平(P＜0．05)。结论 DEX则可能通过上调内源性BDNF mRNA和TrkB mRNA表达，有利于抵御细菌性脑膜炎脑损伤。     

瘦素对四氯化碳所致小鼠急性肝损伤的影响

探讨瘦素对四氯化碳(CCl4)所致的小鼠急性肝损伤是否有加重作用.方法:分别在小鼠腹腔注射CCl4和/或重组小鼠瘦素,测其血清转氨酶(AST和ALT),并取肝组织作病理学观察,用RT-PCR法检测肝组织TGF-β1和α1(Ⅰ)前胶原mRNA表达.结果:与对照组相比,CCl4处理组和CCl4加瘦素处理组小鼠血清转氨酶增高(P均＜0.01);肝脏的炎症和坏死程度明显加重;肝组织TGF-β1和α1(Ⅰ)前胶原的mRNA表达都升高(P＜0.05和P＜0.01).瘦素处理组则无明显变化.但CCl4加瘦素处理组和CCl4处理组相比,各项指标进一步升高(P均＜0.01).结论:瘦素可以明显增加小鼠肝脏急性损伤后的炎症反应和早期纤维的形成,可能是肝纤维化的促进因子.     

益气化痰解毒中药对脂肪肝大鼠脂联素及肝脏瘦素受体mRNA的影响

[目的]探讨益气化痰解毒中药治疗脂肪肝分子生物学机制.[方法]将60只大鼠随机分为正常组与模型组(采用高脂饮食加白酒灌胃建立大鼠脂肪性肝炎模型),模型组所有大鼠再随机分为对照亚组、中药亚组、东宝肝泰亚组.用药4周后,观察并比较模型组及其各亚组与正常组对血清脂联素(APN)、瘦素(leptin)及肝脏瘦素受体(OB-R) mRNA、肿瘤坏死因子-α的影响.[结果]模型组与正常组比较血清leptin浓度升高,APN及肝脏OB-RmRNA表达显著下降(P＜0.01),治疗后中药亚组APN水平明显升高,肝脏OB-R mRNA表达上调,同时血清leptin、肝组织肿瘤坏死因子-α含量下降,明显优于东宝肝泰亚组(P＜0.01).[结论]益气化痰解毒中药通过提高APN,上调肝脏OB-RmRNA的表达,以调节leptin水平达到有效治疗脂肪性肝炎的作用.     

脂多糖受体CD14通过核因子-κB途径参与大鼠急性肝功能衰竭的动态观察

目的 观察CD14、核因子(NF)-κB在D-氨基半乳糖介导的大鼠急性肝功能衰竭模型中的动态变化规律.方法 用D-氨基半乳糖诱导大鼠急性肝功能衰竭模型,在造模后6、12、24、36、48、72、120、168、240 h用鲎试剂检测门静脉脂多糖(LPS),RT-PCR法检测肝脏CD14 mRNA,免疫组织化学检测CD14和NF-κB蛋白表达.实验数据采用两样本t检验及直线相关.结果 CD14蛋白和CD14 mRNA在造模后6 h升高,48 h达高峰(37.13±17.65,0.716±0.089),与健康对照组(2.07±1.84,0.355±0.098)比较,差异有统计学意义(t=-13.236,t=-6.664,P＜0.01),72、120、168和240 h逐渐下降.NF-κB蛋白6 h升高,48 h达高峰(41.97±8.76),与健康对照组(2.33±2.02)比较,差异有统计学意义(t=-24.137,P＜0.01),72 h及以后逐渐下降(P＜0.01).LPS、ALT和AST在6 h有轻度升高,至24 h与健康对照组比较,差异有统计学意义(P＜0.05),48 h达高峰(P＜0.01),72 h及以后逐渐下降(P＜0.01).相关性分析显示,CD14 mRNA、CD14蛋白与ALT、AST、LPS、NF-κB蛋白均有正相关性(r=0.698,P＜0.01).结论 CD14作为LPS的识别受体,可能通过NF-κB途径激活下游细胞因子,引起肝脏损伤.     

趋化因子Fractalkine在急性肝功能衰竭大鼠模型中的变化及意义

目的 探讨不规则趋化因子Fraetalkine(FKN,CX3CLl)在急性肝功能衰竭大鼠模型中的变化及其在肝脏炎性损伤中的作用.方法 SD大鼠分为健康对照组6只,模型组36只.D氨基半乳糖(D-Gal)诱导大鼠急性肝功能衰竭模型,造模后分别在12、24、48、72、120和168 h等6个时间点取大鼠血及肝脏标本,RT-PCR法检测肝组织中FKN mRNA、核因子(NF)-kB mRNA的变化.计量资料组间比较用t检验,相关性检验用Pearson直线相关分析.结果 造模后12 h,大鼠血ALT、AST值明显升高,分别为(208.3±43.5)U/L和(375.25:117.3)U/L,显著高于正常组的(31.8±2.9)U/L和(90.8±3.1)U/L,差异有统计学意义(t值分别为-9.912和-5.935,P＜0.01),72 h达高峰.造模后12 h,FKN mRNA为0.086±0.009,高于正常组的0.044±0.009,差异有统计学意义(t=-7.999,P＜0.01),72 h达高峰,为0.333±0.033,120 h则明显下降.NF-kB在正常大鼠肝组织中有少量表达,模型组随着时间推移,NF-kB水平逐渐升高,72 h达高峰,为0.583±0.101(t=-12.607,P＜0.01).FKN与NF-kB呈正相关(r=0.760,P＜0.01).结论 FKN在急性肝功能衰竭大鼠中的表达是肝损伤的重要因素,有可能为急性肝功能衰竭的治疗提供一个新的切入点.     

MMP2与细粒棘球蚴感染小鼠肝纤维化研究

目的通过检测细粒棘球蚴感染小鼠肝脏中MMP2及其mRNA表达水平的变化,探讨其在肝纤维化过程中的作用。方法收集感染细粒棘球蚴病羊肝脏,取肝脏囊泡中的原头蚴处理后接种实验组小鼠肝脏,建立细粒棘球蚴病动物模型。于感染后2、8、30、90和180d各取8只小鼠处死,无菌收集肝脏,采用RT-PCR检测MMP2mRNA,采用免疫组织化学法检测MMP2在肝脏中的表达。实验设假手术对照组。结果与对照组比较,实验组小鼠肝脏MMP2mRNA水平在感染后2d达高峰(t2d=6.566,P0.01),总体呈现早期高表达,晚期下降的趋势。免疫组化检查实验小鼠肝脏MMP2表达趋势与RT-PCR的结果一致。结论小鼠感染细粒棘球蚴早期肝脏MMP2高表达,中、晚期低表达,这可能是导致细粒棘球蚴小鼠发生肝脏纤维化的机制之一。     

增龄及去卵巢对C57BL/6J小鼠护骨素表达的影响

目的观察增龄和去卵巢状态下小鼠血清护骨素(OPG)水平和骨组织中OPG表达的变化。方法4组C57BL/6J小鼠,每组15只：成年雄鼠组(6月龄),老年雄鼠组(20月龄)；假手术雌鼠组和去卵巢雌鼠组鼠在9月龄时分别行假手术和卵巢切除术,手术6个月后处死小鼠。双能X线检测小鼠骨量变化,实时RT-PCR观察OPG mRNA表达水平,ELISA测定血清中OPG的蛋白水平。结果老年组小鼠和去卵巢组小鼠骨密度明显下降。老年组雄性小鼠血清中OPG水平和成年组[(729±180)ng/Lvs (565±131)ng/L]比较明显上升(P＜0.01)。和假手术组比较(644±140)ng/L,去卵巢组小鼠OPG水平(908±338)ng/L也升高(P＜0.05)。老年组雄鼠骨中OPG mRNA的表达拷贝数为成年组的43.75%(P＜0.05)。与假手术比较,去卵巢组雌鼠骨中OPG mRNA的表达拷贝数下降了75%(P＜0.01)。结论增龄以及去卵巢状态下．小鼠血清中OPG以及骨中OPG mRNA表达水平均有明显变化。     

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.