多聚酶链式反应随机引物扩增的DNA多态指模技术在幽门螺杆菌菌株鉴别中的应用

本利用多聚酶链式反应随机引物扩增的DNA(RAPD)多态指模技术研究其在HP菌株鉴别的应用。对47例株HP进行RAPD多态指模分析。结果发现：①40株由不同病人分离的HP菌株有不同的DNA指模；②同一菌株多次培养后其DNA指模无改变；③HP根除疗法后3个月4例、6个月1例(分离的HP菌株分与治疗前相同，提示可能是HP复发；④HP根除疗法后6个月及24个月各1例)与治疗前分离的菌株DNA指模不同．提示可能是HP再感染．结果提示RAPD多态指模技术对HP菌株的鉴别有价值。     

江苏省三起甲型副伤寒爆发菌株与散发菌株的基因型及药物敏感性比较

目的 比较江苏省三地区连续发生的甲型副伤寒爆发与散发菌株在分子水平的联系,及其药物敏感性.方法 采用随机扩增多态性DNA(RAPD)分析方法,对爆发及散发菌株进行基因分型,采用K-B法对两类菌株进行药敏测定.结果 31株菌呈四个基因型图谱,爆发与散发菌株中基因Ⅰ型皆占优势,爆发株对链霉素、氨苄西林、头孢拉啶的耐药率从4.6%～25.9%.结论 RAPD方法能给予表型相同的甲型副伤寒菌株进行基因分型,基因Ⅰ型为江苏省甲型副伤寒的优势菌株.     

呼吸道感染铜绿假单胞菌随机扩增DNA多态性分型研究

用随机扩增DNA多态性分析技术（RAPD）对31株铜绿假单胞菌进行分型．结果显示31株铜绿假单胞菌共有9种不同的基因型,其中基因A型17株（占54．8％）。提示基因A型为优势菌株,且多重耐药。经对RAPD技术优化,本研究建立了较稳定的检测系统。     

嗜肺军团菌血清I型菌株随机扩增多态性DNA指纹图谱研究

目的采用随机扩增多态性DNA(RAPD)方法研究我国58株嗜肺军团菌血清1型(Lp1)菌株的基因型特征。方法58株嗜肺军团菌血清1型菌株来自北京和深圳市的集中空调冷却塔水和温泉水,随机引物采用5’-CGGCGGCG-GCGG-3’序列,分析Lp1型军团菌RAPD基因型和温度、pH值的关系。结果58株Lp1型菌株共分为17个RAPD基因型。与集中空调冷却塔水Lp1型菌株相比,温泉水分离的菌株基因型具有较高的多态性。深圳市29株集中空调冷却塔水分离的Lp1菌株,共呈现出3个RAPD基因型,北京市24株来自温泉水的Lp1菌株分为12个RAPD型,5株来自集中空调冷却塔水的Lp1型菌株分为两个RAPD型。温泉水Lp1菌株的基因型差异和水温度和pH值无明显的关联。结论RAPD分型方法可用于我国Lp1菌株的基因分型,不同地区和来源的Lp1型军团菌菌株具有特征性的RAPD基因型,温泉水中分离的Lp1菌株较集中空调冷却塔水分离菌株基因型呈现较高的基因多态性。     

金黄色葡萄球菌随机扩增多态性DNA指纹图与药敏谱对医院感染管理的指导意义

目的 探讨金黄色葡萄球菌(SA)药敏谱型对医院感染管理的指导意义。方法 通过随机扩增多态性DNA(RAPD)技术分型76株SA，并归纳出药敏谱型与RAPD型的对应关系。结果 药敏谱型35种，RAPD分型50种。二者呈单一对应关系的有22种，呈一对二关系的有12种，呈一对三关系的有1种。结论 依据SA药敏谱型可对SA进行分型，从而指导医院感染管理。     

幽门螺杆菌感染个体中菌株基因多态性的研究

幽门螺杆菌(H.pylori)感染的临床结局与宿主易感性、环境因素和菌株的基因多态性相关。了解感染个体中菌株的基因多态性，有助于阐明H.pylori感染的影响因素、致病机制和治疗后复发等问题。目的：研究同-H.pylori感染个体中的菌株是否存在基因多态性。方法：分别从14例临床H.pylori感染者的胃窦活检标本中分离、培养H.pylori,从每例菌株中分离出10个单克隆菌株。从分离自另7例患者胃窦和胃体活检标本的配对菌株中各分离出1个单克隆菌株。采用随机扩增多态性DNA(RAPD)指纹分析方法检测H.pylori菌株的基因多态性。结果：分离自不同感染个体的H.pylori菌株的RAPD指纹图有显著差异。大多数来自同一胃活检部位或同一感染个体的单克隆菌株的RAPD指纹图相同或十分相似，仅少数单克隆菌株的指纹图存在差异。结论：不同H.pylori感染个体所携带菌株的基因型存在显著多态性，同一个体携带的H.pylori菌株基因型相对均一。     

流感嗜血杆菌耐药菌株基因分型的初步研究

目的建立随机扩增DNA多态性(RAPD)反应体系,对海口市流感嗜血杆菌(Hi)耐药菌株进行基因分型。方法对临床分离的62株Hi进行药物敏感试验。同时采用RAPD技术对耐药菌株进行基因分型,并对分型结果进行聚类分析。结果 62株Hi中有23株耐药,其中7株(第10～16菌株)对氨苄西林、氯霉素、四环素、复方新诺明耐药,2株(第17～18菌株)对四环素、复方新诺明耐药,1株(第19菌株)对复方新诺明、阿奇霉素耐药,9株(第1～9菌株)对氨苄西林耐药,4株(第20～23株)只对阿奇霉素耐药。采用引物(5′-ACGTATCTGC-3′)构建的RAPD-PCR反应体系检测Hi基因,扩增出15条带,大小在300～2 000bp之间。所构建的亲缘关系聚类图中,第1～9菌株聚成一簇,第10～16菌株聚成一簇,第17～18菌株聚成一簇,第19菌株为一簇,第20～23菌株聚成一簇。结论采用RAPD-PCR反应体系可对Hi耐药菌株进行基因分型。     

中国鼠疫菌随机引物扩增多态性指纹图谱

目的 对中国不同鼠疫自然疫源地鼠疫菌染色体的分析和比较发现其近缘关系进而从分子水平进行分型。方法 用随机扩增多态性DNA(RAPD)技术。结果 第1条引物将中国的鼠疫菌分为11个类型，每个类型都有一定的条带规律和地理分布范围。第2条引物将我国的鼠疫菌分为5个类型。结论 随机扩增多态性DNA(RAPD)可以从分子水平用于我国鼠疫菌的分型。     

小肠结肠炎耶尔森氏菌的随机扩增多态性DNA的分析

分别对从福建、浙江、沈阳等地分离到的18个血清型的115株小肠结肠炎耶尔森氏菌用随机扩增多态性DNA（RAPD）方法进行了分析，将其分为22个型别。结果提示RAPD方法比血清分型方法的优点是对于追踪传染源有更高的价值。     

广州市儿童A族链球菌T型连续7年调查

目的　调查广州市学龄儿童咽部携带A族链球菌 (GAS)T型在不同时期的流行规律和特点。方法　从1988年 9月至 1994年 7月 ,分别在广州市越秀区 (城市 ) ,广州市番禺区大石镇 (农村 )采用随机整群抽样的方法在小学3～ 5年级的小学生中抽取 2～ 3个班学生作为研究对象 ,研究时间分别为 1988年 9月至 1989年 8月、1990年 5月至1991年 4月、1992年 9月至 1993年 7月、1993年 8月至1994年 7月共 4个学年度 ,跨度达 7年的长期观察和研究 ;研究期内每个月对每个受试者进行一次咽拭子培养 ,将分离出的 β 溶血性链球菌进行杆菌肽试验 ,兰菲分组 ,T分型和OF分型。结果　共分离出 2 5 30株GAS ,T型分布结果依序为T不可分型 (UT)占 17 6 8%、T6 11 2 3%、T4 9 2 9%、T17 79%、T115 6 1%、T8/ 2 5 /IMP195 4 2 %和T5 / 2 7/ 4 44 98%等 7个占总菌株 5 %以上的主导菌型 ;城乡两地菌型差异较大 (P <0 0 0 1) ;城乡两地T型在不同年份变化较大(P <0 0 0 1) ;4 2 4株GAS作OF试验 ,结果OF阴性菌株占6 5 8%。结论　广州市 1988年 9月至 1994年 7月之间 ,学龄儿童咽部流行的GAST型主导菌株依序为T6、4、1、11、8/2 5 /IMPl9和 5 / 2 5 / 4 4为主 ,城乡两地流行的主导菌型差异较大 ,不同年份GAST型分布也存在很大差异     

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.     

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.     

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.     

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.