我国首次发现的暗纹节菱孢所致皮肤真菌病的毒理实验及病理分析

从患者皮损处皮屑及活检组织中分离出的暗纹节菱孢Ａｒｔｈｒｉｎｉｕｍｐｈａｅｏｓｐｅｍｕｍ（Ｃｏｒｄａ）Ｅｌｌｉｓ菌株制成混悬菌液,用兔,豚鼠及小鼠（ＬＡＣＡ）进行动物毒理实验,表皮直接涂抹法接种,四周后未见皮肤损害,划破表皮涂沫法及皮下注射法接种,两周后实验动物的皮肤都出现浸润性斑块,小结节,皮下脓肿及脱毛的症状,对照组动物正常,再用腹腔注射法接种,小鼠两周后死亡,病解所见,腹膜,肠系膜,腹壁及肝     

我国首次发现暗孢节菱孢所致皮肤真菌病——一株新的条件致病性真菌

本文报告暗孢节菱孢(又名球形阜孢霉)所致皮肤真菌病。此菌是我国首次发现的条件致病真菌。患者头部发生多处皮下结节及萎缩性秃发。皮损活检可见毛囊和皮脂腺严重破坏。肉芽肿内有褐色孢子和分隔菌丝,尤以皮下脂肪浸润明显。多次从皮损外皮屑内检出圆形的厚壁孢子。在沙氏琼脂培养基上,37℃培养生长良好。菌落初呈淡黄色,渐变为橙色。小培养镜检时(放大400倍),可见菌丝分枝有隔(1—2μm)。单细胞性小分生孢子呈球形,晶体形等,光滑发亮呈褐色。有的孢子中央有一黑色纵条带。扫描电镜下发现球形或卵形分生孢子,将暗孢节菱孢制成的菌液接种于实验动物皮肤,2周后出现斑块、小结节、小脓肿和脱毛等症状,与患者临床症状相同。对照组动物正常。动物皮损活检所见病理损害与患者皮损病理损害相同,证实了此菌的致病性。     

暗孢节菱孢菌引起面部感染1例及相关研究

目的报道暗孢节菱孢菌(Arthrinium phaeospermum)引起的面部皮肤感染1例并进行相关真菌学研究。方法患者女,29岁,两颊对称性浸润性红斑数年。皮损直接镜检、真菌培养,经形态观察、分生鉴定菌种、并进行体外药敏试验及利用豚鼠致病性实验重现组织病理特征。结果直接镜检未见明显菌丝,体外培养后为单一菌落,经棉兰染色、小培养、扫描电镜、分子生物学均鉴定为暗孢节菱孢菌(Arthrinium phaeospermum),均可见典型特征。体外药敏试验提示,该菌对卡泊芬净、米卡芬净、泊沙康唑、雷夫康唑、伏立康唑、特比萘芬敏感,对伊曲康唑、氟康唑耐药。豚鼠致病性实验中,对照组和免疫抑制组豚鼠皮下注射处大体可见脓肿,体表涂抹处大体表现红斑、丘疹、脱屑,免疫抑制组症状持续较久。皮下注射处较体表涂抹处有明显的组织病理学改变,可见多核巨细胞包裹菌丝或孢子,伴组织细胞、中性粒细胞、淋巴细胞浸润。结论该患者为罕见的暗孢节菱孢菌(Arthrinium phaeospermum)引起的皮肤真菌感染。免疫功能对该菌感染所致疾病的临床过程可能有影响。     

乙型肝炎疫苗-卡介苗联合疫苗动物安全性试验研究

为评价乙型肝炎疫苗-卡介苗联合疫苗的安全性,分别给豚鼠皮下注射、小鼠皮内注射乙型肝炎疫苗-卡介苗联合疫苗或卡介苗,豚鼠每2周称体重一次,观察6周,解剖检查其病理变化;小鼠也进行病理检查。结果乙型肝炎疫苗-卡介苗联合疫苗接种组和卡介苗接种组的豚鼠体重均增加,疫苗注射局部及各脏器病理改变相似;小鼠接种局部皮肤病理改变也未见差异。结论为乙肝表面抗原(HBsAg)没有促使卡介苗毒力增强,乙型肝炎疫苗-卡介苗联合疫苗接种实验鼠是安全的。     

皮内与腹腔接种不同来源菌株对实验性小鼠孢子丝菌病的影响

目的将不同来源的申克孢子丝菌菌株接种于免疫抑制处理后的小鼠皮内及腹腔,造成其实验性感染,观察其感染情况,研究不同菌株及不同接种途径对小鼠孢子丝菌病的影响。方法分别在小鼠皮内和腹腔注射孢子丝菌菌悬液0.1ml,约含1×107个孢子,观察3周。观察结束时处死小鼠取其血、脑、肺、肝、脾、肾、肠系膜、睾丸、腹膜进行真菌培养和组织病理检查。结果皮内和腹腔接种不同来源菌株的小鼠各器官感染率各不相同。播散株皮内接种组、固定株腹腔接种组、播散株腹腔接种组的各器官平均感染率分别为16.05%、18.89%、58.73%。播散株腹腔接种组与其他二组之间差异均有显著性,脾和睾丸是最易受累的器官。结论不同来源菌株可能毒力不同,播散型菌株具有更强的侵袭性。脾脏可能在宿主抗孢子丝菌感染中具有重要作用。     

探讨接种时间和剂量对小鼠肺炎支原体肺炎模型建立的影响

目的 用不同时间和剂量建立BALB/c小鼠的肺炎支原体肺部感染模型,探索小鼠急性肺炎支原体感染的过程.方法 BALB/c小鼠随机分为Ⅰ、Ⅱ两组,Ⅰ组在0、1、2 d滴鼻接种肺炎支原体菌液3次,Ⅱ组在0 d滴鼻接种小剂量肺炎支原体菌液1次后于8、9 d再接种和Ⅰ组相同的肺炎支原体菌液2次,均设立不同剂量、批次的生理盐水对照组.全部实验动物在3～18 d内分批处死.所有小鼠均取肺组织做病理切片.以组织病理学评分来确定小鼠的肺部炎症反应程度.取肺组织匀浆及肺泡灌洗液(BALF)进行肺炎支原体培养做病原学检测.结果 接种后实验动物全部存活无死亡.实验组动物均出现不同程度的肺炎支原体肺炎样病理改变,组织病理学评分为1.5～14.3分,分别于第5、6天和第11天达到最高,平均分为4.9分;Ⅰ组实验组动物多呈轻、中度改变,Ⅱ组实验组动物多呈中、重度改变.所有实验组动物肺组织匀浆及BALF的肺炎支原体培养在接种后1月内先后出现阳性结果.对照组动物未出现明显肺部炎症改变,组织病理学评分为0～1分,肺炎支原体培养为阴性.结论 成功建立BALB/c小鼠的肺部肺炎支原体感染模型;组织病理学评分方法可用以评价肺部炎症反应的严重程度;不同接种时间和剂量所致的病变程度不同.     

不同品系小鼠对实验感染念珠状链杆菌敏感性的观察

本文通过念珠状链杆菌（Ｓｔｒｅｐｔｏｂａｃｉｌｕｓｍｏｎｉｌｉｆｏｒｍｉｓ,Ｓ．ｍ．）实验感染昆明、Ｃ５７ＢＬ／６Ｊ、ＢＡＬＢ／ｃ、ＩＣＲ、ＮＩＨ、ＤＢＡ共６个品系小鼠,观察其对Ｓ．ｍ．敏感性的差异。其中昆明、Ｃ５７ＢＬ／６Ｊ两个品系在腹腔接种后表现出明显的临床、病理改变。昆明小鼠发病率和死亡率分别为９２％和８０％;Ｃ５７ＢＬ／６Ｊ分别是８０％和１２％。昆明小鼠较之Ｃ５７ＢＬ／６Ｊ小鼠起病急、病情严重,多数动物死于感染的急性期。其余品系的发病率为：ＮＩＨ８％、ＢＡＬＢ／ｃ和ＤＢＡ４％,没有动物死亡;ＩＣＲ在接种后无任何临床病理改变。在实验感染后临床病理改变方面,昆明小鼠和Ｃ５７ＢＬ／６Ｊ小鼠间有较大不同。昆明小鼠以末梢血管淤血、四肢尾部水肿、关节炎、截瘫、腹泻为主,而Ｃ５７ＢＬ／６Ｊ小鼠则以注射部位和其它部位皮下脓肿、化脓性关节炎为主。本研究提示,中国昆明小鼠对Ｓ．ｍ．敏感性最高,可以将其作为“哨兵动物”用于实验大鼠Ｓ．ｍ．的常规监测;不同品系小鼠不仅对实验感染Ｓ．ｍ．的敏感性不同,而且表现出不同的临床病理改变。     

超声和CT在诊断腹膜后冷脓肿中的应用

目的：评估超声和ＣＴ诊断腹膜后冷脓仲的价值。材料与方法：用超声和ＣＴ对４１例腹膜后冷脓肿作了检查,观察了腹膜后冷脓肿的形态、大小、位置、内部回声或密度情况、活动度以及对周围组织、脏器的影响。结果：肾周腹膜后的大脓肿,其形态呈前后径小,上下径大的不规则梭形,小脓肿或髂窝处的脓肿多呈类圆形。超声的内部回声有低回声、无回声、等回声和强回声,ＣＴ为脓肿内见斑点及沙砾样钙化灶,增强扫描后的病灶更为清楚。所有     

Ⅱ型糖尿病并发结核病小鼠模型的建立及评价

目的建立Ⅱ型糖尿病并发结核病小鼠模型,对其肺组织病变过程进行评价。方法高脂高糖饲料喂养C57BL/6J小鼠,腹腔注射STZ,建立Ⅱ型糖尿病小鼠模型。采用滴鼻的方式将结核分枝杆菌H37Rv接种至糖尿病小鼠体内,建立糖尿病并发结核病小鼠模型。分别于感染后的1~8周、10周及12周解剖小鼠,肉眼观察小鼠肺脏病变情况,活菌菌落计数,肺组织行HE及抗酸染色,镜下观察病理学改变。结果肉眼观察发现,随着感染时间的增长肺部感染情况逐渐加重,至第8周时病灶达到全肺的80%,第10周时病变范围开始缩小;感染8周后HE染色肺组织出现典型的肉芽肿结构,至第10周部分病变肺组织出现修复现象;抗酸染色可见结核分枝杆菌。结论从大体病变、菌落计数及病理改变等方面对小鼠模型进行评价,发现糖尿病并发结核病小鼠模型较单一的结核病小鼠模型具有病变出现早,病程进展快等特点,与临床患者的某些阶段表现类似,因此实验构建的糖尿病并发结核病小鼠模型基本成功。     

结核分枝杆菌感染小鼠的脾脏和肺脏组织荷菌量与病理变化

目的分析结核急性感染小鼠脾脏和肺脏的组织荷菌量以及病理的动态变化。方法以3×105CFU结核分枝杆菌标准株H37Rv尾静脉途径感染雌性C57BL/6J小鼠,测定感染后1 d、1周、2周、3周、4周、6周和8周肺脏及脾脏组织的荷菌量,脾脏和肺脏组织经HE染色分析病理变化。结果感染动物的脾脏荷菌量在前3周逐步提高,在4～8周脾脏荷菌量维持稳定,肺脏组织在2～8周组织荷菌量逐步升高。病理分析显示动物感染3周后肺脏和脾脏出现肉芽肿,在6～8周病理损伤加重。结论小鼠经尾静脉感染高剂量结核分枝杆菌在8周前表现为急性感染状态,适用于抗结核药物的体内药效学评价。     

Vertebrate safety of Clostridium bifermentans serovar malaysia, a new larvicidal agent for vector control.

The safety of bacterial cells of Clostridium bifermentans serovar malaysia, which is highly toxic to mosquito larvae, was tested on mice, guinea pigs, rabbits, and goldfish. Inoculations of at least 1 x 10(8) cells per animal by routes recommended by World Health Organization (subcutaneous, percutaneous, inhalation, force-feeding, intraperitoneal, intravenous) and tests of subacute toxicity, anaphylactic shock, persistence in heart blood, and virulence by successive passages, were performed on mice, guinea pigs, or both. Growth was monitored for 1 mo before necropsy. Ocular irritation and skin scarification were tested with rabbits. C. bifermentans serovar malaysia did not induce any mortality or abnormal reactions in mammals at a dose 1,000 times higher than the level established by W.H.O. for the demonstration of safety. Bacterial cells are not toxic to goldfish at a dose 1,000 times higher than the LD50 for the target-mosquito larvae. We conclude that C. bifermentans serovar malaysia bacterial cells are safe for laboratory mammals and goldfish.     

Allergenicity of Mycobacterial Ribosomal and Ribonucleic Acid Preparations in Mice and Guinea Pigs

Guinea pigs were injected subcutaneously with mycobacterial ribosomal fraction incorporated in Freund's incomplete adjuvant and tested 6 and 12 weeks later by the intradermal injection of 0.5 μg (25 TU) of Purified Protein Derivative. No evidence of delayed-type hypersensitivity could be detected in these animals, although large necrotic reactions were obtained in guinea pigs sensitized with living, attenuated mycobacterial cells. Mice also were vaccinated by the intraperitoneal injection of mycobacterial ribosomal fraction or ribonucleic acid (RNA) and tested for sensitivity to tuberculin at various subsequent times. No evidence of true tuberculin hypersensitivity could be detected at any time, although what appeared to be small Arthus type reactions were seen in mice given the largest vaccinating doses. Attempts to recall tuberculin sensitivity in vaccinated mice by the intravenous injection, 4 weeks after vaccination of living cells, of either the virulent or attenuated mycobacterial strains were unsuccessful. Instead, when the virulent cells were injected, a suppression of footpad reactivity was noted in animals made sensitive to tuberculin by the previous intraperitoneal injection of viable attenuated mycobacterial cells. Both guinea pigs and mice, vaccinated as described above, were also skin tested or footpad tested, respectively, with 2 μg of the ribosomal fraction or RNA used for vaccination. No evidence of true tuberculin hypersensitivity could be obtained; instead, in guinea pig skin very small dermonecrotic areas were noted, and in mice swelling and redness of the footpad occurred to an equal extent in both vaccinated and nonvaccinated mice. The possible role of tuberculin hypersensitivity in acquired immunity to tuberculosis is discussed, and the conclusion is reached that its part, if any, is minor.     

Changes in responses of UVB irradiated skin of brownish guinea pigs with aging

It is known that skin often shows irregular pigmentation during aging, which is frequently associated with hyperpigmentation. Many studies have utilized brownish A1 guinea pigs to investigate the pathogenesis of ultraviolet B (UVB)-induced skin pigmentation, however, responses associated with aging following UVB irradiation have not been elucidated. To characterize those responses, dorsal skin of A1 guinea pigs from 14-weeks to 5-yr old were investigated. The minimal erythema dose was found to increase with aging. Further, in pigmentation induced by UVB radiation, skin brightness (DeltaL*-value) decreased equally in both the 14-week old (young) group and in the 3-yr old (old) group of guinea pigs. The DeltaL*-value recovered in the young group from 21 d after UVB irradiation, whereas no such recovery was seen in the old group. In addition, the amount of melanin and the number of melanocytes returned near pre-irradiation levels in the young group, while they remained high in the old group. Our results therefore demonstrate for the first time that skin responses following UVB irradiation change with aging in A1 guinea pigs.     

Comparison of adjuvants with Leishmania antigens in a guinea pig model to induce delayed-type hypersensitivity responses.

BACKGROUND AND PURPOSE: Guinea pigs have been a traditional model for studies of delayed-type hypersensitivity. They are the natural host of Leishmania enriettii and have been experimentally infected with other species of Leishmania. They have been used as a skin-test model to screen potential antigens for use in diagnostic tests for Leishmania. Use of complete Freund's adjuvant (CFA), along with whole promastigote Leishmania antigen, was necessary to sensitize guinea pigs to invoke a sufficient cell-mediated immune response. However, use of CFA has come under scrutiny by Animal Care and Use Committees due to the pathologic changes associated with its use. METHODS: Thirty-two specific-pathogen-free male Hartley guinea pigs were inoculated with Leishmania antigens alone or mixed with one of three adjuvants (CFA, TiterMax, and liposomes), and were skin tested 2 weeks later. RESULTS: For the Leishmania antigens tested, guinea pigs that received liposomes as an adjuvant had skin-test responses comparable to those of guinea pigs that received CFA. TiterMax was also tested, but cellular responses at antigen test sites were poor. CONCLUSIONS: Liposomes can be used in this model as a safe, effective adjuvant.     

结核疫苗保护力评价用感染菌液的制备和保藏研究

实验研究中对结合分枝杆菌感染菌液进行活菌数量与毒力的稳定性观察。以活菌计数与感染豚鼠后的肝、脾、肺病变指数为评判指标进行观察,结果显示低温保藏菌液在32个月内,其活菌数量处于6.7～18×105CFU/ml之间,感染豚鼠的肝、脾、肺病变指数处于42～51之间。说明低温保藏菌液具有很好的稳定性,可作为结核分枝杆菌感染豚鼠模型标准化用菌液。     

Toxicity of Pasteurella tularensis killed by ionizing radiation

Approximately 2 x 10(11) viable Pasteurella tularensis cells per ml, contained in suspensions, were killed by exposure to 10(6) r of gamma-radiation. When injected intraperitoneally into mice, the irradiated suspensions initially contained about 10 ld(50) per ml, and immunized mice against challenge with fully virulent strains of P. tularensis. Toxicity and immunizing activity of the suspensions decreased significantly within a few days at 5 C. Mice were protected against the toxin by immune serum or by prior injection of endotoxin of Escherichia coli. Cortisone did not protect against the newly prepared suspension, but was effective against the aged suspension. Lethal doses of newly prepared suspension for guinea pigs and rabbits were approximately 0.5 ml and 2 ml, respectively. Cortisone protected rabbits, but not guinea pigs, against lethal challenge. Pyrogenic effects resembling those shown by endotoxin-containing suspensions were demonstrated in rabbits. The results suggested that two toxins are responsible for the toxicity of irradiated suspensions of P. tularensis: one labile and associated with the immunizing activity of the suspension, the other more stable and resembling classical endotoxin.     

Pigmentation in intrinsically aged skin of A1 guinea pigs

It is known that skin often shows irregular pigmentation during aging which is frequently associated with hyperpigmentation. Many studies have utilized brownish A1 guinea pigs to investigate the pathogenesis of ultraviolet (UV)-induced skin pigmentation, however, changes associated with intrinsic aging in A1 guinea pig skin have not been documented. To characterize such changes, skin from the dorsal and neck areas of 20-week, 1-, 2-, 3- and 5-yr-old guinea pigs was examined. Skin color was measured using a colorimeter, and biopsy specimens were stained with Masson-Fontana, L-3,4-dihydroxyphenylalanine (DOPA), and antibodies against KIT (ACK-45), gp100 (HMB-45) and S-100 proteins. The L* value of skin color decreased with aging and melanin deposits increased in the epidermis. Further, DOPA+, gp100+ and S-100+ melanocytes increased, indicating that the number of melanocytes had increased with age, whereas KIT+ melanocytes did not increase in dorsal skin and actually decreased in neck skin with aging. Further, rippled pigmented areas appeared in the neck skin of the 3-yr-old animals, and in the dorsal and neck skin of 5-yr-old guinea pigs in the absence of UV irradiation. Melanocytes were distributed uniformly in younger skin, whereas they were clustered in older skin. UV irradiation caused an increase in the number of melanocytes, although they were not clustered. These results are the first to provide evidence that pigmentation is induced in the skin of intrinsically aged A1 guinea pigs in the absence of UV irradiation, a process that differs from that elicited by UV irradiation.