利福布汀抗结核分枝杆菌活性及其耐药性与rpoB基因突变关系的探讨

目的 比较利福平和利福布汀的体外抗MTB活性,分析其交叉耐药性,评价利福平和利福布汀耐药性与rpoB突变的关系.方法 采用微量平板Alamar Blue检测(MABA)法,检测利福平和利福布汀对MTB临床分离菌株168株的抗菌活性,并对其rpoB基因核心区进行DNA序列测定.两组间基因突变率的比较采用x2检验.结果 66株利福平敏感菌株均对利福布汀敏感( MIC≤0.125 mg/L),未检测到rpoB基因耐药决定区存在突变;102株利福平耐药菌株中有76株同时对利福布汀耐药( MIC ＞0.5 mg/L),交叉耐药率为74.5％(76/102);26株MIC≤4 mg/L的利福平耐药菌株仍对利福布汀敏感(MIC≤0.5 mg/L);516、526和531位点的氨基酸改变与利福平和利福布汀耐药均有关,而511和533位点的氨基酸改变可能仅与利福平耐药有关,利福布汀耐药菌株的rpoB基因突变率(92.1％,70/76)显著高于利福布汀敏感菌株(23.9％,22/92),差异有统计学意义(x2=78.12,P＜0.05).结论 对利福平低度耐药(MIC≤4 mg/L)的MTB仍对利福布汀敏感.MTB的rpoB基因突变可用于利福平耐药性检测,且对检测利福布汀的耐药性也有参考意义.     

利福平和利福布汀对结核分枝杆菌交叉耐药率的初步探讨

目的通过对结核分枝杆菌MIC的测定,初步探讨利福平和利福布汀两药间的交叉耐药率。方法利用Alarmar Blue法(Alamar blue susceptibility test,MABA)检测116株结核分枝杆菌临床分离株对利福平和利福布汀的体外MIC。结果 36株利福平敏感株均对利福布汀敏感;80株利福平耐药株中有58株对利福布汀也同时耐药(对利福布汀的MIC>0.5 ug/ml),利福平与利福布汀间的交叉耐药率为72.5%(58/80);同时对利福平和利福布汀耐药的菌株对两种药物的耐药程度呈现正相关。结论利福布汀较利福平有更好的体外抗菌活性。对于利福平耐药而利福布汀敏感的患者仍可选择利福布汀作为联合化疗的有效药物之一。     

结核分枝杆菌对利福喷汀与利福平交叉耐药的实验研究

目的 观察利福喷汀对结核分枝杆菌的杀菌效力及其与利福平的交叉耐药性，探讨利福喷汀对结核分枝杆菌的实验室耐药界限，为临床应用利福喷汀治疗结核病，包括治疗对利福平耐药的结核病提供依据。方法 用7H9液体培养基、苏通液体培养基、改良罗氏固体培养基测定利福喷汀和利福平对结核分枝杆菌标准株（H37Rv)及临床分离的利福平敏感株、利福平耐药株的最低抑菌浓度(Minimal Inhibitory Concentration，MIC)。结果 在7H9液体培养基上对临床分离的19株利福平敏感株分别测定利福平和利福喷汀的MIC，有近80%利福平敏感株利福平的MIC≥0．32μg/ml，而利福喷汀的MIC多分布在0．02～0．32μg/ml之间(占84．2％)：无论标准株H37Rv、利福平敏感株(19株)或是利福平耐药株(45株)，利福喷汀对结核分枝杆菌的MIC普遍比利福平低2～4倍：利福喷汀敏感株和耐药株的MIC差别较大,在MIC为5～ 10μg／ml处能最大限度地区分敏感菌和耐药菌。结论 利福喷汀与利福平存在着交叉耐药。但是利福喷汀比利福平有更强的杀菌效力，部分结核分枝杆菌对利福平达到临床耐药时，仍未达到利福喷汀临床耐药。对利福平耐药的部分结核分枝杆菌对利福喷汀仍具有一定程度的敏感性,提示临床上对利福平耐药的结核病患者使用利福喷汀可能有一定效果；实验结果对利福喷汀临界耐药界限进行了初步筛查,为利福喷汀常规药敏试验的开展提供了基本试验数据。     

氯法齐明对不同耐药类型结核分枝杆菌的体外抑菌活性研究

目的 评价氯法齐明对不同耐药类型的MTB临床分离菌株的体外抑菌活性,为临床应用提供依据.方法 应用微孔板观察法,测定氯法齐明对MTB标准株H37Rv及临床分离敏感、单耐药、多耐药、耐多药和广泛耐药菌株各15株的MIC,并将氯法齐明对不同耐药类型的MTB菌株体外活性与异烟肼、利福平、阿米卡星、卷曲霉素和氧氟沙星进行比较.结果 氯法齐明对敏感菌株的MIC(0.06～4.00mg/L)高于异烟肼(0.06～0.25 mg/L)和利福平(0.06～0.25 mg/L),与氧氟沙星(0.06～2.00mg/L)、阿米卡星(0.06～4.00mg/L)和卷曲霉素(0.50～4.00mg/L)相似;氯法齐明对单耐药菌株的MIC(0.03～4.00mg/L)与异烟肼(0.06～0.25 mg/L)和利福平(0.06～0.25 mg/L)相似,低于氧氟沙星(0.25～8.00mg/L)、阿米卡星(0.06～4.00mg/L)和卷曲霉素(0.50～8.00mg/L);氯法齐明对耐多药菌株的MIC(0.06～8.00mg/L)与阿米卡星(0.25～8.00mg/L)相似,优于氧氟沙星(0.125～8.00mg/L)和卷曲霉素(0.50～8.00mg/L);氯法齐明对广泛耐药菌株的MIC(0.03～8.00mg/L)明显优于其他药物.结论 氯法齐明对MTB菌株,尤其是耐多药和广泛耐药MTB菌株,均有较好的体外抑菌活性.

Abstract：

Objective To study the in vitro antituberculous activities of clofazimine (CLF) to different drug-resistant types of Mycobacterium tuberculosis.Methods The minimal inhibitory concentration (MIC) of CLF and isoniazid (INH), rifampicin (RFP), ofloxacin (OFLX), amikacin (AK), and capreomycin (CPM) against sensitive, single-drug resistant (SDR), poly-drug resistant (PDR), multi-drug resistant (MDR), and extensive-drug resistant (XDR) Mycobacterium tuberculosis strains isolated clinically were determined by microplate assays.Results The MICs of CLF for sensitive, SDR, PDR, MDR and XDR strains of clinically isolated Mycobacterium tuberculosis were 0.06 -4.00 mg/L, 0.03 -4.00 mg/L, 0.06 -8.00 mg/L, 0.06 - 8.00 mg/L, 0.03 - 8.00 mg/L.For the sensitive group, the MIC of CLF ( 0.06 -4.00 mg/L) was higher than that of INH (0.06 -0.25 mg/L) and RFP (0.06 -0.25 mg/L), while there was no significant difference among OFLX (0.06 -2.00 mg/L), AK (0.06 -4.00 mg/L), and CPM (0.50 -4.00 mg/L).For the single-drug resistant group, there was no significant difference among CLF (0.03-4.00 mg/L), INH (0.06-0.25 mg/L), and RFP (0.06-0.25 mg/L), but the MIC of CLF was lower than that of OFLX ( 0.25 - 8.00 mg/L), AK ( 0.06 - 4.00 mg/L ), and CPM ( 0.50 - 8.00 mg/L).For the MDR group, there was no significant difference between CLF (0.06 -8.00 mg/L) and AK (0.25 - 8.00 mg/L), but the MIC of CLF was lower than that of OFLX (0.125 - 8.00 mg/L) and CPM (0.50 -8.O0 mg/L).For the XDR group, the MIC of CLF (0.03 -8.00 mg/L) was lower than that of others.Conclusion CLF showed good in vitro activity against Mycobacterium tuberculosis, especially MDR and XDR strains.     

结核分枝杆菌对利福喷汀与利福平交叉耐药的实验研究

目的 观察利福喷汀对结核分枝杆菌的杀菌效力及其与利福平的交叉耐药性，探讨利福喷汀对结核分枝杆菌的实验室耐药界限，为临床应用利福喷汀治疗结核病，包括治疗对利福平耐药的结核病提供依据。方法 用7H9液体培养基、苏通液体培养基、改良罗氏固体培养基测定利福喷汀和利福平对结核分枝杆菌标准株(H37Rv)及临床分离的利福平敏感株、利福平耐药株的最低抑菌浓度(MIC)。结果 在7H9液体培养基上对临床分离的19株利福平敏感株分别测定利福平和利福喷汀的MIC，有80％利福平敏感株利福平的MIC≥0．32μg／ml，而利福喷汀的MIC多分布在0．02～0．32μg／ml之间(占84％)；无论标准株H37Rv、利福平敏感株(19株)或是利福平耐药株(45株)，利福喷汀对结核分枝杆菌的MIC普遍比利福平低2～4倍；利福喷汀敏感株和耐药株的MIC差别较大，在MIC为5～10μg／ml处能最大限度地区分敏感菌和耐药菌。结论 利福喷汀与利福平存在着交叉耐药，但利福喷汀比利福平有更强的杀菌效力，部分结核分枝杆菌对利福平达到临床耐药时，仍未达到利福喷汀临床耐药，对利福平耐药的部分结核分枝杆菌对利福喷汀仍具有一定程度的敏感性，提示临床上对利福平耐药的结核病患者使用利福喷汀可能有一定效果；实验结果对利福喷汀临界耐药界限进行了初步筛查，为利福喷汀常规药敏试验的开展提供了基本试验数据。     

广州地区719株淋球菌对壮观霉素 环丙沙星的敏感性测定

目的 了解广州地区淋球菌对壮观霉素、环丙沙星的敏感性。方法 以琼脂稀释法测定719株淋球菌对壮观霉素、环丙沙星的最小抑菌浓度(MIC)。结果 壮观霉素MIC范围为2～128mg／L，MIC50和MIC90分别为16mg／L和32mg／L。719株淋球菌中有718株(99．86％)对壮观毒素敏感，6年间未发现耐药菌株。环丙沙星MIC范围0．002-32mg／L，3．06％菌株对环丙沙星敏感，22．95％为低度敏感，73．99％为耐药。结论 壮观霉素作为治疗淋病的一线药物对淋球菌临床分离株仍相当有效，氟喹诺酮类药物因高耐药率已不再适宜作为治疗淋球菌感染的首选药。     

结核分枝杆菌临床株对利福霉素类药物的耐药特点分析

目的比较分析临床标本结核分枝杆菌(Mycobacterium tuberculosis,Mtb)对利福霉素类药物的耐药特点,为含利福霉素类药物的抗结核治疗方案的改进提供细菌学基础。方法用绝对浓度法检测470份临床标本中Mtb的耐药情况。结果 180份培养阳性的标本中,41份对利福霉素类耐药,对利福平和利福喷汀的耐药水平高度一致,对二者的耐药水平显著高于对利福布汀的耐药水平,对低浓度和高浓度利福布汀耐药的数量差异显著。结论临床标本中的Mtb对利福平和利福喷汀的耐药水平表现出高度一致性,提示利福喷汀不适于作为利福平耐药的后备药物,利福布汀可以作为利福平的后备药物,但服用剂量可能影响治疗效果。     

达托霉素对2679株革兰阳性球菌外抗菌活性的研究

目的 研究达托霉素等抗菌药物对2679株革兰阳球菌的体外抗菌活性.方法 收集2010年1月-2011年12月9个城市17家教学医院临床分离的2679株非重复革兰阳性球菌.采用微量肉汤稀释法测定的达托霉素最低抑菌浓度(MIC),用琼脂稀释法测定其他抗菌药物的MIC值,用WHONET5.6软件进行药敏数据统计分析.结果 耐甲氧西林金黄色葡萄球菌(MRSA)和耐甲氧西林凝固酶阴性葡萄球菌(MRSCoN)检出率分别为45.8％和84.2％,MRSA对复方磺胺甲(噁)唑和氯霉素的敏感率分别为93.1％和85.5％,对红霉素、四环素、克林霉素和利福平的敏感率分别为13.8％、26.6％、63.2％、50.0％,对达托霉素、万古霉素和利奈唑胺的敏感率均为100.0％,MRSCoN对达托霉素、万古霉素和利奈唑胺的敏感率均为100.0％,达托霉素对于MRSA和MRSCoN的MIC50和MIC90均为0.5 mg/L.513株肠球菌对于高水平庆大霉素耐药率为56.9％,氯霉素和四环素的敏感率分别为76.0％和44.1％,对替加环素和达托霉素敏感率均达100.0％,达托霉素对于其中17株耐万古霉素肠球菌(V RE)的MIC50和MIC90均为2 mg/L.肺炎链球菌和β-溶血链球菌对于达托霉素的敏感率均为100.0％,按口服青霉素折点判读,青霉素不敏感的肺炎链球菌(PNSSP)的比例为63.1％.达托霉素对于PNSSP的MIC50和MIC90分别是0.125 mg/L和0.25 mg/L.达托霉素对于β-溶血链球菌MIC50和MIC90分别是0.008 mg/L和0.032 mg/L.结论 达托霉素对临床常见革兰阳性球菌具有较好的抗菌活性,包括多重耐药菌,是治疗革兰阳性菌特别是耐药菌感染的很好选择.     

耐多药结核病快速诊断试剂盒检测耐药结核分枝杆菌的评价

目的 评价用耐多药结核病快速诊断(GenoType MTBDRplus)试剂盒检测耐利福平、异炳肼MTB的效果.方法 选取75株MTB临床分离株,采用耐多药结核病快速诊断试剂盒检测其中的耐药MTB,评价其敏感度和特异度及可重复性,并与常规的药物敏感性试验方法相比较.结果 耐多药结核病快速诊断试剂盒对上海地区的MTB临床分离株中耐利福平和耐异烟肼菌株的敏感度分别为98.0%和86.5%,对敏感菌株的特异度为100%.而在重复性试验中,耐多药结核病快速诊断试剂盒多次结果完全一致,重复性达到100%.结论 耐多药结核病快速诊断作为快速检测耐药MTB的试剂盒,对耐利福平和耐异烟肼菌株具有较高的敏感度和特异度,在上海地区有较好的应用前景.     

氟喹诺酮类药物对动物源MRSA和MSSA的防耐药变异浓度比较

目的比较4种氟喹诺酮类药物对动物源性MRSA和MSSA的耐药突变体的选择能力。方法采用琼脂平板稀释法分别测定环丙沙星、左氧沙星、加替沙星和吉米沙星对MRSA和MSSA的MIC值和MPC值,并计算MIC90和MPC90。结果 4种氟喹诺酮药物对85株动物源性MSSA的MPC90值分别为4mg/L、2mg/L、0.5mg/L、0.5mg/L,加替沙星的MPC90/MIC90比值最小;对21株动物源性MRSA的MPC90值分别为128mg/L、64mg/L、16mg/L、8mg/L,吉米沙星的MPC90/MIC90比值最小。结论对动物源性MSSA,加替沙星防耐药突变能力优于其它,加替沙星和吉米沙星单药能有效限制耐药突变株的选择,而左氧沙星和环丙沙星则易选择耐药菌株。对动物源性MRSA,4种药物单药给药均不能有效限制耐药突变株的选择。     

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.