结核分支杆菌利福平耐药基因突变的研究

目的了解我国结核分支杆菌耐利福平（ＲＦＰ）分离株ｒｐｏＢ基因突变情况，建立快速检测结核分支杆菌耐药基因型的分子药敏试验方法。方法通过聚合酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）和ＰＣＲ－直接测序法（ＰＣＲ－ＤＳ）分析５０株结核分支杆菌临床分离株的ｒｐｏＢ基因。结果３株结核分支杆菌药物敏感株和２株非ＲＦＰ耐药株中，仅１株ｒｐｏＢＳＳＣＰ图谱异常的药物敏感株出现５３１位密码子ＴＣＧ→ＴＴＧ突变。４５株ＲＦＰ耐药株中，１０株ＳＳＣＰ和ＤＳ分析均未见ｒｐｏＢ突变，３５株ＳＳＣＰ和ＤＳ分析异常，其中１４株为５３１位密码子ＴＣＧ→ＴＴＧ或ＴＧＧ或ＴＡＣ突变，１４株为５２６位ＣＡＣ→ＴＡＣ或ＧＡＣ或ＣＣＣ或ＣＴＣ或ＧＴＣ突变，２株为５１６位ＧＡＣ→ＧＴＣ或ＴＡＣ突变，２株为５１６位和５２６位及５１５位和５１６位密码子双点突变，３株ｒｐｏＢ序列与结核分支杆菌不同。结论大多数结核分支杆菌耐ＲＦＰ是由于其ｒｐｏＢ基因突变所致，采用ＰＣＲ－ＳＳＣＰ和ＰＣＲ－ＤＳ方法可快速测定结核分支杆菌ＲＦＰ耐药基因型     

临床分离多重耐药假单胞菌耐药机制的研究

为了解假单胞菌对亚胺培能（ＩＭＰ）、头孢他啶（ＣＴＺ）、头孢哌酮（ＣＰＺ）和氨曲南（ＡＺ）的耐药机制。以绿脓杆菌标准株ＰＡＯ１和对照株ＴＮＰＯＯ４为对照，应用ＳＤＳ－ＰＡＧＥ外膜蛋白图谱和β－内酰胺酶测定方法，研究了临床分离的两株多重耐药绿脓杆菌和一株ＩＭＰ耐药荧光假单胞菌。结果表明：两株绿脓杆菌分离株均缺失了外膜蛋白ＯｐｒＤ＿２和外膜蛋白Ｅ（ＯｐｒＥ），并出现了４４．７ＫＤ的未知蛋白带；两株菌均产生了较高水平的β－内酰胺酶，其酶活性分别为０．６８１Ｕ／ｍｇ和０．６ｌ０Ｕ／ｍｇ，经ＩＭＰ诱导，两株菌产酶量未见显著增加；其β－内酰胺酶对上述药物的相对水解率均小于１０％。提示该两株绿脓杆菌对上述药物的多重耐药机制可能是ＯｐｒＤ＿２、ＯｐｒＥ的缺失和大量β－内酰胺酶的共同作用结果；其中对ＩＭＰ耐药的主要原因可能是ＯｐｒＤ＿２丢失。另一株ＩＭＰ耐药荧光假单胞菌出现了ＯｐｒＤ＿２含量的减少。     

急性髓系白血病多药耐药表达与白血病克隆生长的关系

多药耐药（ＭＤＲ）及白血病细胞高度增殖能力所产生的再生耐药,是急性髓系白血病（ＡＭＬ）化疗失败的重要原因。我们采用植物血凝素——白细胞条件培养液（ＰＨＡ－ＬＣＭ）为生长刺激因子,体外培养白血病祖细胞（Ｌ－ＣＦＵ）。同时使用ＪＳＢ－１单克隆抗体免疫组化ＡＢＣ法检测ＡＭＬ病人骨髓细胞中Ｐ１７０表达,以探讨两者间关系及其临床意义。１　材料与方法１．１　研究对象　２１例ＡＭＬ均经细胞形态学、免疫学分型确诊。ＦＡＢ分型Ｍ１６例,Ｍ２５例,Ｍ３５例,Ｍ４１例,ＣＭＬ急变４例。其中初治１８例,复发２例,难治性…     

多重耐药大肠杆菌外膜屏障机制的研究

目的研究临床分离的多重耐药大肠杆菌耐药的外膜屏障机制。方法用尿素－ＳＤＳ－ＰＡＧＥ法对临床分离的大肠杆菌外膜孔蛋白进行分析，并用抽滤法测定大肠杆菌细胞内四环素浓度。结果临床分离的多重耐药大肠杆菌Ｒ３、Ｒ４、Ｒ２８、Ｒ３０外膜孔蛋白中均缺失ＯｍｐＦ，Ｒ３、Ｒ３０中同时缺失ＯｍｐＣ，Ｒ１２缺失ＯｍｐＡ。敏感菌株中均不缺失ＯｍｐＦ，但敏感株Ｓ２２缺失ＯｍｐＡ和ＯｍｐＣ。ＬＭ２１８（ＯｍｐＦ标准缺失株）、Ｒ２８细胞中稳态四环素浓度分别为敏感株Ｓ３６的８０％和４０％。结论外膜通透性的改变是大肠杆菌产生多重耐药的机制之一     

浙江省耐甲氧西林金黄色葡萄球菌流行状况及耐药性

对我省各地区１９９１年６月至１９９２年１２月间收集的１７０株临床分离的金黄色葡萄球菌的研究表明：在我省同样存在耐甲氧西林金黄色葡萄球菌（ＭＲＳＡ）的流行，在１７０株菌株中共检出了ＭＲＳＡ４４株，占同期金黄色葡萄球菌的２５．９％，且在大多数地区都检测到了ＭＲＳＡ菌株；对其耐药性进行研究发现，ＭＲＳＡ菌株对大多数抗生素的耐药率都比较高，对有的抗生素的耐药率在９０％以上，耐药程度明显高于甲氧西林敏感的金黄色葡萄球菌（ＭＳＳＡ），但对万古霉素全部敏感。     

结核分支杆菌链霉素耐药基因的检测

目的了解结核分支杆菌链霉素（Ｓｍ）耐药基因突变情况，建立快速检测耐药突变株的方法。方法通过聚合酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）分析了２２株结核分支杆菌临床分离株的ｒｐｓＬ基因，并应用ＰＣＲ－限制性片段长度多态性（ＰＣＲ－ＲＦＬＰ）进一步分析其突变的部位和性质。结果以Ｈ３７Ｒｖ标准株为对照，５株敏感株的ｒｐｓＬ基因未见ＳＳＣＰ泳动异常、有ＭｂｏⅡ酶切位点；１３株耐Ｓｍ株中，１１株（８６％）有ｒｐｓＬ基因泳动异常、无ＭｂｏⅡ酶切位点；４株耐其它抗结核药物株中，也有１株ＳＳＣＰ泳动异常、并无ＭｂｏⅡ酶切位点。结论大多数结核分支杆菌Ｓｍ耐药分离株有ｒｐｓＬ突变，而突变位点大多位于第４３位密码子。应用ＰＣＲ－ＳＳＣＰ和ＰＣＲ－ＲＦＬＰ可能成为快速检测结核分支杆菌耐药突变株的新方法。     

浙江省耐甲氧西林金黄色葡萄球菌流行况及耐药性

对我省各地区１９９１年６月至１９９２年１２月间收集的１７０株临床分离的金黄色葡萄球菌的研究表明：在我省两样存在耐甲氧西林金黄色葡萄球菌（ＭＲＳＡ）的流行，在１７０株菌株中共检出了ＭＲＳＡ４４株，占同时期金黄色葡萄球菌的２５．９％，且在大多数地区都检测到了ＭＲＳＡ菌株；对其耐药性进行研究发现：ＭＲＳＡ菌株对大多数抗生素的耐药率都比较高，对有的抗生素的耐药率在９０％以上，耐药程度明显高于甲氧西林敏感的     

甲氧西林耐药金黄色葡萄球菌耐药性的研究

甲氧西林耐药金黄色葡萄球菌（ＭＲＳＡ）感染发生率有逐年增加趋势。为了解ＭＲＳＡ耐药特点及合理选用抗菌药物，我们测定了１８种抗菌药物对临床分离金黄色葡萄球菌（ＳＡ）的最低抑菌浓度（ＭＩＣ）、受试菌的β－内酰胺酶及凝固酶分型。结果表明ＭＲＳＡ占临床分离ＳＡ的５０．７％，其中产β－内酰胺酶菌株占８５．１％，ＭＲＳＡ凝固酶均为ＩＩ型。ＭＲＳＡ对多数β－内酰胺类及某些氨基糖甙类抗生素呈多重耐药性。万古霉素、阿贝卡星对ＭＲＳＡ显示最强的抗菌活性；阿米卡星、妥磺沙星、氧氟沙星、亚胺培南亦显示较强抗菌活性，提示上述药物可用于ＭＲＳＡ所致严重感染。     

碘杂环化合物IHC－72对心肌部分除极和触发活动的影响

用常规玻璃微电极技术，观察３，６－（二甲氨基）－二苯骈碘杂环依地酸盐（ＩＨＣ－７２）对豚鼠右０００室乳头肌细胞膜部分除极引发的慢反应动作电位（ＳＡＰ）、哇巴因引发的延迟后除极（ＤＡＤ）及触发活动（ＴＡ）的影响。２．５４μＭＩＨＣ－７２对ＳＡＰ各参数均无影响；２５．４，１０１．６μＭＩＨＣ－７２分别使动作电位幅值（ＡＰＡ）减少７．４％和１９．３％，０相最大上升速率（Ｖ＿（ｍａｘ））减少２．５％和３９．１％，复极１００％的动作电位时程（ＡＰＤ＿（１００））缩短７．１％和４１％。有效不应期（ＥＲＰ）相对延长４．９％和９ｌ．３％，静息膜电位（ＲＭＰ）不受影响。浓度为２５．４μＭＩＨＣ－７２对复极５０％的动作电位时程（ＡＰＤ＿（５０））无影响，而浓度为１０１．６μＭ时能使ＡＰＤ＿（５０）缩短４２．９％，且能抑制由哇巴因诱发的ＤＡＤ及ＴＡ。结果表明：ＩＨＣ－７２对慢钙内流有直接抑制作用；其抑制触发性心律失常的机理与钙拮抗有关。     

临床分离耐氟喹诺酮金黄色区区球菌的耐药机制研究

为了解临床分离金黄色葡萄球菌（金葡萄）对氟喹诺酮类药物的耐药机制。应用聚合酶链反应－限制性片段长度多生（ＰＣＲ－ＲＦＬＰ）分析和ＰＣＲ－边构象多太分析（ＳＳＣＰ）技术及利血平逆转实验，分别检测３１株临床分离耐环丙沙星金谋略 株ｇｙｒＡ基因的突变及ｎｏｒＡ基因的表达。结果表明，３１菌中２２株检出ｇｒｙＡ基因８４位点突变，有２８株在得血平逆转实验中对环丙沙星和诺氟沙星的ＭＩＣ同时降低，表明存在 ｏｒＡ     

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.     

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.     

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.     

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.