小鼠肝多房棘球蚴中血管新生与病程发展的相关性研究

目的初步探讨小鼠肝多房棘球蚴中血管新生与疾病发生发展的相关性。方法将60只6～8周龄的C57BL/6雌性小鼠随机分成实验组和对照组,每组30只。小鼠经5%水合氯醛溶液麻醉后开腹,实验组小鼠肝脏注射多房棘球蚴原头节混悬液100μl (约400个原头节),对照组注射等量PBS,无菌缝合关腹。分别于感染后30、 60、 90、 120 d,肝脏灌注法观察病变组织血管分布和新生情况;尾静脉采血, ELISA检测小鼠血清中血管内皮生长因子A (VEGFA)的表达;实验组取棘球蚴组织和棘球蚴周围肝组织,对照组取相应肝组织,制备石蜡切片,苏木素-伊红(HE)染色观察病理组织学改变;免疫组织化学染色法观察不同感染时间、不同部位肝脏组织中VEGFA、 CD34和CD31的表达情况。结果肝脏灌注法结果显示,随着感染时间的延长,实验组小鼠肝脏棘球蚴组织逐渐增大,周围可见明显的血管分布。ELISA检测结果显示,感染后30、 60、 90和120 d,实验组小鼠血清中VEGFA浓度分别为269.00、 420.62、 539.00和271.73 pg/ml,差异有统计学意义(P 0.01),且均高于相应对照组的VEGFA浓度(194.00、 173.00、 234.00和127.00 pg/ml)(P 0.05)。HE染色结果显示,随着感染时间的延长,实验组小鼠棘球蚴组织体积逐渐增大,并伴有嗜酸性粒细胞浸润、结核样肉芽组织形成和纤维组织增生,病变内部区域可见原头节和钙化灶。免疫组织化学结果显示,感染后棘球蚴组织中均可见VEGFA、 CD34和CD31不同程度的表达,阳性染色定位于棘球蚴组织内皮细胞。感染后30、 60、 90和120 d,棘球蚴组织中VEGFA免疫组织化学评分分别为(1.60±0.52)、(3.10±0.87)、(4.80±1.32)和(2.40±1.07)分,其中感染后30 d与感染后60 d、 90 d比较差异均有统计学意义(P 0.05, P 0.01);棘球蚴组织与棘球蚴周围组织(VEGFA评分均为0)和对照组(VEGFA评分均为0)比较差异均有统计学意义(P 0.01)。感染后30、 60、 90和120 d,棘球蚴组织中微血管密度(CD34-MVD)分别为(38.70±11.06)/HP、(65.50±8.46)/HP、(109.90±9.40)/HP和(56.86±6.64)/HP,差异有统计学意义(P 0.001),且均高于棘球蚴周围组织和对照组(P 0.01)。棘球蚴组织中CD31-MVD分别为(19.80±3.12)/HP、(30.70±2.50)/HP、(47.90±4.77)/HP和(31.10±3.84)/HP,差异有统计学意义(P 0.01),且均低于Em周围组织和对照组(P 0.001)。结论多房棘球蚴浸润性生长中伴有血管新生, VEGFA的分泌刺激了血管新生,新生血管促进棘球蚴的生长。     

泡球蚴感染C57BL/6小鼠肝脏CD34表达的动态变化

目的 观察泡球蚴(Em)感染C57BL/6小鼠肝脏CD34微血管密度(MVD)的动态变化。方法 将40只C57BL/6小鼠随机分为实验组和对照组,实验组通过肉眼肝叶穿刺感染泡球蚴(Em),对照组通过注射等量PBS。分别于感染后30 d、60 d、90 d及120 d后取小鼠肝脏组织,苏木素-伊红(HE)染色观察肝脏的病理组织学改变,免疫组织化学染色观察泡球蚴感染后CD34-MVD的动态改变。结果 实验组泡球蚴组织各时间点中CD34-MVD分别为(39.10±11.84)/HP、(66.80±11.08)/HP、(111.20±8.00)/HP和(56.14±7.12)/HP,实验组泡球蚴周围肝组织分别为 (1.14±0.82)/HP、(1.56±0.92)/HP、(2.32±1.43)/HP和(1.38±0.82)/HP ;对照组分别为(1.00±0.94)/HP、(1.30±1.06)/HP、(2.00±1.15)/HP和(1.10±0.87)/HP。实验组与对照组比较,差异均有统计学意义(P<0.01);实验组泡球蚴组织内比较,差异有统计学意义(F=94.63,P<0.001);实验组泡球蚴周围肝组织内比较,差异无统计学意义(F=3.24,P>0.05);对照组内比较,差异无统计学意义(F=1.98,P>0.05)。结论 泡球蚴浸润性生长过程中可能伴随着血管新生,血管新生可能为其生长重要的机制之一。     

实验性慢性氟中毒大鼠致密胶原组织的光镜及电镜变化

本文观察了慢性氟中毒大鼠致密胶原组织——肌腱的光镜变化,发现肌腱内胶原组织成熟老化不够,胶原纤维较细,胶原蛋白形成较少。电镜改变则见胶原微纤维走行紊乱,粗细不均,甚至溶解断裂。对氟中毒时胶原纤维形态改变的机制及防治措施进行了讨论。     

膝骨关节炎前交叉韧带Ⅰ型和Ⅲ型胶原纤维定量分析及组织学相关研究

目的 探讨老年膝骨关节炎（OA）前交叉韧带（ACL）退变时Ⅰ、Ⅲ型胶原纤维的分布、含量与组织学改变特点。方法 选择膝关节OA患者全膝关节置换术中切取的ACL27例（29膝）和正常膝关节新鲜ACL标本5例，采用苦味酸-天狼猩红染色观察并分析ACL中Ⅰ、Ⅲ型胶原纤维的分布、含量和比例改变。HE染色观察ACL的组织学变化。结果 苦味酸-天狼猩红染色发现，正常ACL胶原排列整齐，偏振光下呈横纹肌样排列，主要由Ⅰ、Ⅲ型胶原构成，Ⅰ型胶原粗大，含量较多，在韧带内呈弥漫性分布；Ⅲ型较少，多分布在各束韧带的内部。OA患者的ACL胶原排列紊乱，Ⅲ型胶原含量增多，Ⅰ型胶原明显减少。HE染色组织学观察到OA患者ACL松散，有不同程度的纤维结缔组织变性，表现为不均匀的纤维细胞数目增多或减少，可见黏液变性、软骨样化生、囊性变及玻璃样变性，并有淋巴细胞浸润。结论 苦味酸-天狼猩红染色可以应用于观察并计算ACL中Ⅰ、Ⅲ型胶原的分布与含量的变化。膝关节OA伴有ACL退变时Ⅰ、Ⅲ型胶原纤维的分布与含量有改变．ACL退变与OA具有相关性。     

环氧合酶-2在四氯化碳诱导肝硬化大鼠肝窦毛细血管化形成中的作用

目的 观察环氧合酶-2(COX-2)在实验性肝硬化大鼠肝窦毛细血管化形成中的作用.方法 腹腔注射CCl4每周2次共8周诱导雄性SD大鼠肝硬化模型.将SD大鼠分成3组:正常对照组(n=10)、模型对照组(n=15)和罗非昔布治疗组(10 mg·kg-1·d-1,n=15).光镜下观察肝组织标本,电镜观察肝窦超微结构改变.用Western印迹和免疫组化法检测基底膜蛋白主要成分层粘连蛋白(LN)和Ⅳ型胶原,同时通过Ⅷ因子相关抗原(vWF)免疫组化标记微血管牛成密度.结果 与模型对照组相比,罗非昔布干预治疗能减少肝纤维化面积(分别为30.7±8.9和23.5±6.5,P<0.05).光镜及电镜提示,在模型对照组可见肝窦内皮细胞窗孔减少、缩小,有完整的基底膜形成,Disse腔隙内有大量的胶原纤维沉积,罗非昔布组上述病变有所减轻.随着肝硬化的形成,肝组织微血管密度明显升高,罗非昔布组肝组织微血管密度(6.4±0.7)较模型对照组(11.3±1.6)明显降低(P<0.01).肝硬化时肝组织表达Ⅳ型胶原和LN蛋白明显增加(分别为3.8±0.4和3.7±0.5),罗非昔布能降低Ⅳ犁胶原和LN的表达(分别为3.0±0.5和3.0±0.5;与模型对照组相比两者均为P相似文献   

颈椎黄韧带光电镜结构和胶原含量的增龄变化及其意义

目的：研究椎黄韧带光电镜结构和胶原含量的增龄变化及其临床意义,方法对比观察胎儿,婴幼儿、成人,老年人及颈椎病患者颈椎黄韧带的光电镜结构,并用Ｗｏｅｓｓｎｅｒ法测定其胶原含量。结果胎儿组黄韧带中弹性蛋白稀少,外周无或仅有极少量的短微原纤维。婴幼儿组弹性蛋白增多,外周已出现短微原纤维,结果与成人组接近。老年人组弹力纤维减少,胶原纤维增多,且变性,钙化,此改变在患者组更明显,更广泛。从胎儿组,婴幼儿组到     

冠脉微循环障碍参与慢性心力衰竭大鼠心肌重构

目的探讨冠脉微循环障碍在慢性心力衰竭(CHF)大鼠心肌重构中的地位与作用。方法Wistar大鼠随机分为实验组、对照组。实验组皮下注射异丙肾上腺素(340mg/kg),对照组皮下注射生理盐水0.5ml,连续2次,间隔24h。第2次注射结束3个月后测定血液动力学及左心室质量/体质量。左心室心肌行HE染色,Masson胶原染色,透射电镜检查,凝集素组织化学染色显现毛细血管。结果与对照组比较,实验组左心室收缩压、左心室压力上升和下降最大速率降低,左心室舒张末压、左心室质量/体质量升高。与对照组比较,实验组心内膜下心肌灶性坏死,胶原增加,电镜显示毛细血管痉挛。与对照组比较,实验组心内膜下心肌毛细血管密度[(1972±83)/mm2对(1633±62)/mm2]、心肌细胞密度[(1778±97)/mm2对(1588±87)/mm2]显著降低(P<0.001),毛细血管密度与心肌细胞密度的比值[(1.11±0.08)对(1.03±0.05)]显著降低(P<0.05)。结论冠脉微循环障碍参与CHF大鼠心肌重构,并且是造成CHF大鼠心肌重构的病因之一。     

CVB3m重复感染心肌致心室重构的实验研究

目的探讨CVB3m重复感染致病毒性心肌炎心肌结构的改变。方法用梯度倍增法连续4次经腹腔注射CVB3m病毒液感染昆明鼠，分别于末次感染后10、30、60d处死动物，电镜下观察心肌细胞超微结构的改变，HE染色观察心肌病变程度和分布，VG染色观察心肌纤维增生情况。结果(1)电镜下显示末次感染后10d股丝走行紊乱，线粒体大部分肿胀，少数嵴溶解消失，肌浆网扩张，溶酶体糖原颗粒增多。30d时肌丝紊乱程度减轻，糖原减少，核周有轻度的低电子密度区，线粒体嵴仍有断裂或絮状变，基质密度降低。60d时心肌间质中血管周围有大量纤维素的沉积，胶原纤维合成增多。(2)HE染色显示末次感染后10d心肌损伤严重，30d时仍可见小灶坏死，60d时未检出心肌损伤。(3)VG染色表明末次感染后10d心肌即出现较弱的胶原染色，30d时胶原明显可见，60d时心肌组织的胶原显著增加，心肌组织血管周围胶原面积(RCVA)和心肌胶原容积分数(CVF)均显著增加。结论CVB3m重复感染可导致急性期严重的心肌坏死，慢性期心肌胶原沉积和心肌纤维化。     

重组2型腺相关病毒-转化生长因子β3对实验性肝纤维化大鼠肝脏组织病理学和Ⅰ型胶原表达的影响

目的 通过重组2型腺相关病毒(rAAV2)和基因转导,探讨转化生长因子(TGF)β 3对人鼠肝纤维化的影响. 方法构建并验证rAAV2-TGF 3.实验人鼠随机分为正常对照组、模型组、阴性对照组和TGF β 3干预组.40%的四氯化碳复制大鼠肝纤维化模型.在四氯化碳诱导前l周,将rAAV2 TGF β 3 注入大鼠尾静脉,造模8周后处死人鼠,取肝脏组织做HE染色观察肝脏组织学改变,Masson染色观察胶原纤维分布,免疫组织化学染色观察Ⅰ型胶原的表达,对胶原纤维的阳性面积比和Ⅰ型胶原的平均吸光度值进行半定量分析.多组数据问比较采用单因素的方差分析,若各组总体方差齐同,采用SNK-q检验,若方差不齐,采用Durmett'S T3检验. 结果 HE染色结果显示,rAAV2-TGF β3干预后,肝组织纤维间隔增生减少;Masson染色结果显示,人鼠胶原纤维主要分布在血管和汇管区及狄氏间隙,TGF β 3干预组大鼠肝内的胶原纤维阳性面积比为7.7%±1.5%,明显低于模型组的13.2%±2.2%和阴性对照组的12.3%±1.5%(q值分别为9.456和8.217,P值均<0.01).免疫组织化学染色结果显示,Ⅰ型胶原主要表达在血管和汇管区及狄氏间隙,TGF β 3干预组大鼠肝内的Ⅰ型胶原的表达为0.185±0.033,明显低于模型对照组的0.252±0.042和阴性对照组的0.230±0.029(口值分别为6.228和4.346,P值均<0.01).结论 TGF β 3可明显减轻实验人鼠的肝纤维化程度和组织学损伤,降低Ⅰ型胶原的表达.     

非结核分枝杆菌性淋巴结炎病理学改变的初步研究

目的 探讨非结核分枝杆菌性淋巴结炎的特征性组织病理学改变.方法 建立非结核分枝杆菌性淋巴结炎的实验动物模型,光镜下观察组织病理学改变;对1例临床初步诊断为非结核分枝杆菌性淋巴结结核患者的病理组织标本经多重PCR方法检测筛选后,光镜下观察组织病理学改变.结果 非结核分枝杆菌感染所致的淋巴结炎的病理学改变与我们所熟知的淋巴结结核的病理学改变不完全相同.在动物模型中观察到的非结核分枝杆菌淋巴结炎的特征性病理学改变如下:(1)淋巴结内可见结节状肉芽肿形成,中央呈凝固性坏死,其中可见较多的中性粒细胞及其核碎屑.坏死区周围可见类上皮细胞、淋巴细胞、单核细胞,并可见纤维组织包裹.肉芽肿内外均可见朗汉斯巨细胞;(2)淋巴结内可见匐形性坏死,坏死旱长条状,中央部分为凝固性坏死,其中有大量中性粒细胞及其核碎屑,周围可见类上皮细胞、淋巴细胞及单核细胞,并可见纤维组织包绕;(3)淋巴结内可见星状和星芒状坏死.对初步诊断为非结核分枝杆菌性淋巴结炎的1例石蜡包埋组织标本,光镜下观察淋巴结内可见上皮样肉芽肿,匐行性坏死和星状坏死,坏死中可见大量中性粒细胞及其核碎屑,周围可见类上皮细胞、淋巴细胞、单核细胞、朗汉斯巨细胞;类上皮细胞的细胞核旱极性排列,周围并可见纤维组织或胶原组织包绕.结论 匐形性坏死、星状和星芒状坏夕匕、中央十酪样坏死中可见大量的中性粒细胞和粒细胞细胞核的碎屑、在中央坏死区的周围可见类上皮细胞胞核的极性排列,是非结核分枝杆菌性淋巴结炎的部分特征性组织病理学改变.     

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.