槲皮素-3-O-α-L-吡喃鼠李糖苷对阿霉素致体外心肌细胞损伤的保护作用

目的观察槲皮素-3-O-α-L-吡喃鼠李糖苷对阿霉素(ADR)致心肌细胞损伤的保护作用及机制。方法实验选用出生24 h内的Wistar大鼠6只,雌雄不限,分离心肌细胞。采用纯化培养的心肌细胞建立ADR损伤模型,实验分为6组:空白对照组(正常细胞培养,加入等数量的MEM培养基,不加入任何药物)、二甲基亚砜(DMSO)对照组(加入含DMSO浓度为5μg/ml的MEM培养液)、ADR损伤组(加AOR浓度为1μg/ml的MEM培养液)、槲皮素-3-O-α-L-吡喃鼠李糖苷保护组(分别为低剂量12.5μg/ml、中剂量25μg/ml、高剂量50μg/ml槲皮素-3-O-α-L-吡喃鼠李糖苷保护组)。分组给药后孵育24 h。显微镜下观察心肌细胞形态,MTT法测定心肌细胞存活率,应用试剂盒分别检测样品中乳酸脱氢酶(LDH)、一氧化氮合酶(NOS)、超氧化物歧化酶(SOD)含量。结果与ADR损伤组比较,低、中、高剂量保护组均可使心肌细胞存活率增高(P<0.01),LOH活性降低(P<0.01或P<0.05),NO活性降低(P<0.01),SOD活性增强(P<0.01)。结论 12.5~50μg/ml槲皮素-3-O-α-L-吡喃鼠李糖苷均能减轻ADR对心肌细胞损伤,具有明显的心肌细胞保护作用,其机制可能与其抗氧化作用有关。     

欣力胶囊对阿霉素所致心肌细胞损伤的保护作用

目的观察欣力胶囊对阿霉素(ADR)中毒心肌细胞的保护作用并探讨其机制。方法采用新生SD乳鼠心肌细胞做原代培养,复制ADR损伤心肌细胞模型,测定培养上清液多项生化指标,并运用流式细胞仪检测凋亡细胞。结果ADR(终浓度为10~(-6)M)可致上清液乳酸脱氢酶(LDH)释放量增加(P<0.01),同时细胞超氧化物歧化酶(SOD)活力下降而丙二醛(MDA)含量升高(P<0.01),欣力胶囊(0.025～0.1 mg/mL)可呈浓度依赖性地降低LDH活力,增加细胞SOD活力和降低MDA含量(P<0.05或P<0.01)。流式细胞仪检测欣力胶囊能降低心肌细胞凋亡率,证实了欣力胶囊的保护作用。结论欣力胶囊能够拮抗ADR引起的自由基脂质过氧化,从而保护心肌细胞。     

铁离子螯合剂去铁敏在大鼠心肌细胞原代培养中对心肌细胞的保护作用

目的:在心肌细胞培养液中添加去铁敏,以对抗原代培养操作过程所产生的心肌细胞损伤,并探讨其保护作用. 方法:在常规新生SD乳鼠心肌细胞原代培养液中按照不同浓度添加去铁敏.抗а-肌动蛋白免疫组化法鉴定心肌细胞.心肌细胞分为对照组(不添加去铁敏);5 μmol/L去铁敏组;15 μmol/L去铁敏组;25 μmol/L去铁敏组.MTT法检测心肌细胞存活率、乳酸脱氢酶活性检测、TUNEL法检测心肌细胞凋亡率评价各组心肌细胞的损伤程度、生存率及凋亡情况. 结果:①5μmol/L去铁敏组(69.6±5.4 IU/L)、15 μmol/L去铁敏组(35.5±3.3 IU/L)和25μmol/L去铁敏组(51.0±4.3 IU/L)的乳酸脱氢酶水平均比对照组(76.3±6.1 IU/L)低,差异均有统计学意义(P<0.05～0.01).15 μmol/L去铁敏组乳酸脱氢酶水平明显低于5 μmol/L去铁敏组和25 μmol/L去铁敏组,差异均有统计学意义(P<0.05～0.01).②15 μmol/L去铁敏组(91.28±2.57)%和25 μmol/L去铁敏组(86.03±4.66)%心肌细胞生存率均比对照组(83.13 4±5.26)%高,差异均有统计学意义(P相似文献   

环孢霉素A对乳鼠心肌细胞结构及功能的影响

目的: 观察环孢霉素A(cyclosporine A,CsA)对体外培养的乳鼠心肌细胞结构及功能的影响,探寻CsA诱导心肌细胞损伤的机制。方法: 采用酶消化法体外培养原代乳鼠心肌细胞,用100 ml/L DMEM培养基培养48 h后,随机分为对照组和CsA干预组。CsA干预组根据CsA的终浓度又分为103 μg/L组、104 μg/L组和105 μg/L组。分别于给CsA后24 h,在光镜及电镜下观察心肌细胞形态结构的变化。用MTT比色法检测细胞数量的变化,于给CsA后24 h及48 h,分别用比色法和硫代巴比妥酸（TBA）法检测细胞上清液中乳酸脱氢酶（LDH）及丙二醛（MDA）含量的变化。结果: 光镜下可见CsA干预组心肌细胞的数量减少,形态欠规则,胞浆中可见空泡,跳动频率减低,且随着用药浓度的增大而变化明显。电镜下可见CsA干预组心肌细胞线粒体肿胀、空泡样变,且随着浓度的增大而逐渐加重。MTT比色法检测显示,CsA干预组的细胞数量减少及活力降低（P<0.05）;而LDH及MDA的水平与对照组比明显升高（P<0.05,P<0.01）,且二者随着CsA浓度的增大及时间的延长而差别显著（P<0.05）。结论: CsA对体外培养的心肌细胞有明显的损伤作用,且呈时间和剂量依赖性,可能与其致线粒体损伤及脂质过氧化有关。     

刺囊酸对缺氧复氧诱导心肌细胞凋亡相关基因表达的影响

目的 研究刺囊酸预处理对原代培养的SD乳鼠心肌细胞缺氧复氧损伤诱导的心肌细胞凋亡相关基因Bcl-2、Bax及聚二磷酸腺苷核糖聚合酶(PARP 89 kDa)表达的影响.方法 实验随机分为6组:对照组,缺氧复氧损伤组,缺氧预处理组及低、中、高剂量刺囊酸组(0.5 μmol/L、5μmol/L和50 μmol/L),分别予以缺氧3h后再复氧2h,检测细胞存活率、乳酸脱氢酶(LDH)活性,Western Blot杂交法检测心肌细胞Bcl-2、Bax及PARP(89 kDa)蛋白表达变化.结果 与对照组比较,缺氧复氧损伤组心肌细胞Bcl-2蛋白表达显著低于对照组(P＜0.01),Bax蛋白及PARP(89 kDa)表达显著高于对照组(P＜0.01),细胞存活率明显低于对照组(P＜0.05).与缺氧复氧损伤组比较,不同剂量的刺囊酸预处理能显著提高细胞存活率,降低LDH活性,呈剂量依赖性;中剂量刺囊酸显著增加Bcl-2蛋白表达(P＜0.05),抑制Bax及PARP(89 kDa)蛋白表达(P＜0.05).结论 刺囊酸预处理可通过上调Bcl-2蛋白表达,抑制PARP(89 kDa)及Bax蛋白表达,抑制细胞凋亡,对抗心肌细胞缺氧复氧损伤.     

链脲佐菌素诱导培养皮层神经细胞损伤作用的观察

目的 研究链脲佐茵素(STZ)对原代培养大鼠皮层神经元是否具有损伤作用.方法 常规原代神经细胞培养的方法,培养新生1 d～3 d的wistar大鼠皮层神经元,通过检测CCK-8,乳酸脱氢酶(LDH)释放检测和calccin-AM染色,观察加入不同浓度STZ(1 μmol/L、10μmol/L、100 μmol/L、1 000μmol/L)36 h后对细胞损伤的作用.结果 STZ处理后对原代培养大鼠皮层神经细胞产生明显的损伤作用,包括细胞活力的下降,活细胞数目的 减少,LDH释放的增加并呈现剂量依赖性.结论 STZ对离体培养的神经细胞具有类似脑室注射同样的神经损伤作用.     

大豆异黄酮对过氧化氢诱导大鼠心肌细胞内钙超载的影响

目的观察大豆异黄酮(SI)对H2O2诱导的原代培养大鼠乳鼠心肌细胞内钙超载的拮抗作用。方法采用Wistar大鼠乳鼠体外心肌细胞培养,将心肌细胞分为对照组、H2O2损伤模型组以及H2O2损伤加SI治疗组:SI0.1、1.0、10μg/ml剂量组,预先24h给予SI后使用0.3mmol/LH2O2损伤心肌细胞,以Fluo-3/AM荧光指示剂负载,应用激光共聚焦显微镜技术,分别于加入H2O2后即刻与15min,检测〔Ca2+〕i变化。结果动态监测15min可见对照组心肌细胞内平均荧光强度值较低;H2O2加入后即刻,细胞内荧光光密度值开始增加,15min后细胞内荧光强度和荧光光密度值显著高于对照组(P0.01);H2O2损伤加SI治疗组:预给SI0.1、1.0、10μg/ml三个剂量组〔Ca2+〕i静息荧光强度值、峰值和钙稳态期荧光强度值均较H2O2损伤模型组明显降低(P0.01);同时可见与对照组比较,三个浓度的S.I.均可使心肌细胞内游离钙离子(〔Ca2+〕i)浓度的平均荧光强度值显著降低(P0.01)。结论 SI能显著减轻H2O2诱导的培养心肌细胞内Ca2+超载,且随着药物浓度的逐渐升高,细胞内钙荧光强度反而逐渐下降,呈现剂量依赖性。     

褪黑素对心肌细胞氧化损伤保护作用的实验研究

目的　探讨氧化损伤时褪黑素 (MT)对心肌细胞形态学变化、细胞存活率、自身一氧化氮 (NO)生成的影响 ,以及NO水平与其它氧化指标的关系。方法　分离培养原代心肌细胞 ,分为对照组、H2 O2 处理组、MT组、MT干预组和N 硝基 L 精氨酸 (L NAME)干预组五组进行对照。用外源性H2 O2 造成氧化损伤 ,用生化法检测培养液中乳酸脱氢酶 (LDH)浓度 ,台盼蓝排斥试验检测细胞存活率 ,硫代巴比妥酸法检测细胞中丙二醛 (MDA)含量 ,用试剂盒检测细胞培养液中NO含量。结果　H2 O2 处理组和L NAME干预组中的细胞存活率、NO含量、LDH含量和MDA含量与对照组比较差异有非常显著性 (P <0 .0 1) ,H2 O2 处理组四项均比对照组明显增高 ,而L NAME干预组中NO含量比对照组明显低。MT干预组与L NAME干预组比较LDH含量和MDA含量明显低 (P <0 .0 1) ,与H2 O2 组加入MT干预后 ,培养液中NO和LDH及MDA含量明显降低 (P <0 .0 1)。结论　心肌细胞在受到H2 O2 损伤时 ,心肌细胞自身产生的NO增加 ,并且在一定程度上参与了对心肌细胞的氧化损伤 ;MT不仅能抑制LDH和MDA的升高 ,还能抑制NO的产生。     

依达拉奉保护H9c2心肌细胞对抗化学性低氧引起的损伤

目的探讨新型的自由基清除剂依达拉奉(EDA)能否保护H9c2心肌细胞对抗化学性低氧引起的损伤。方法应用化学性低氧模拟剂氯化钴(CoCl2)处理H9c2心肌细胞以建立化学性低氧损伤模型。应用CCK-8比色法检测细胞存活率;Hoechst 33258核染色法检测凋亡细胞的形态学及数量变化;双氯荧光素(DCFH-DA)染色荧光显微镜照像测定细胞内活性氧(ROS)水平;JC-1染色荧光显微镜照像检测线粒体膜电位(MMP)。结果应用100～1000μmol/L CoCl2处理H9c2心肌细胞24 h,呈浓度依赖性地降低细胞存活率;在12～36 h范围内,800μmol/L CoCl2呈时间依赖性地抑制细胞存活率;10～40μmol/L EDA或500～2000μmol/L N-乙酰半胱氨酸(NAC,为ROS的清除剂)预处理H9c2心肌细胞1 h呈浓度依赖性地对抗CoCl2对细胞存活率的抑制作用;在800μmol/LCoCl2处理H9c2心肌细胞前1 h,应用40μmol/L EDA预处理细胞不仅可明显的抑制CoCl2诱导的细胞内ROS生成增多,还能抑制CoCl2的致细胞凋亡作用及MMP的损伤作用。结论 EDA能保护心肌细胞对抗CoCl2诱导的损伤作用,此心肌细胞保护作用可能与其抗氧化作用及保护MMP有关。     

白介素33对乳鼠心肌细胞乏氧/复氧损伤时胞内活性氧自由基生成的影响

目的探讨白介素33(IL-33)对乳鼠心肌细胞乏氧/复氧损伤的保护作用及胞内活性氧自由基(ROS)生成的影响。方法分离培养SD乳鼠心肌细胞,将原代培养的心肌细胞随机分为单纯对照(C)组、加药对照(rIL-33 100 ng/ml,C1)组、乏氧/复氧(A/R)组、A/R+rIL-33(1、10、100ng/ml)组,乏氧培养3 h后,复氧2 h。免疫组化鉴定心肌细胞;MTT法检测细胞存活率;ELISA试剂盒检测乳酸脱氢酶(LDH)漏出率;流式细胞仪检测细胞凋亡(AV-PI双标法)、胞内ROS生成量(ROS-DHE荧光探针)。结果与C组相比,A/R组心肌细胞存活率明显下降(P<0.01),LDH漏出显著增高(P<0.01),凋亡率及胞内ROS含量显著增加(P<0.01);与A/R组相比,IL-33治疗组剂量依赖性地提高细胞存活率(P<0.01),降低LDH漏出率、细胞凋亡率(P<0.01)及胞内ROS含量(P=0.03);加药对照组与C组相比,心肌细胞存活率、凋亡率、胞内ROS含量差异无统计学意义。结论 IL-33在心肌乏氧/复氧损伤过程中可以剂量依赖性地发挥抗细胞凋亡作用,其保护作用机制可能与增强心肌抗氧化能力,减少胞内ROS生成有关。     

Three layer functional model and energy exchange concept of aging process

Relying on a certain degree of abstraction, we can propose that no particular distinction exists between animate or living matter and inanimate matter. While focusing attention on some specifics, the dividing line between the two can be drawn. The most apparent distinction is in the level of structural and functional organization with the dissimilar streams of ‘energy flow’ between the observed entity and the surrounding environment. In essence, living matter is created from inanimate matter which is organized to contain internal intense energy processes and maintain lower intensity energy exchange processes with the environment. Taking internal and external energy processes into account, we contend in this paper that living matter can be referred to as matter of dissipative structure, with this structure assumed to be a common quality of all living creatures and living matter in general. Interruption of internal energy conversion processes and terminating the controlled energy exchange with the environment leads to degeneration of dissipative structure and reduction of the same to inanimate matter, (gas, liquid and/or solid inanimate substances), and ultimately what can be called ‘death.’ This concept of what we call dissipative nature can be extended from living organisms to social groups of animals, to mankind. An analogy based on the organization of matter provides a basis for a functional model of living entities. The models relies on the parallels among the three central structures of any cell (nucleus, cytoplasm and outer membrane) and the human body (central organs, body fluids along with the connective tissues, and external skin integument). This three-part structural organization may be observed almost universally in nature. It can be observed from the atomic structure to the planetary and intergalactic organizations. This similarity is corroborated by the membrane theory applied to living organisms. According to the energy nature of living matter and the proposed functional model, the decreased integrity of a human body's external envelope membrane is a first cause of the structural degradation and aging of the entire organism. The aging process than progresses externally to internally, as in single cell organisms, suggesting that much of the efforts towards the restoration and maintenance of the mechanisms responsible for structural development should be focused accordingly, on the membrane, i.e., the skin. Numerous reports indicate that all parts of the human body, like: bones, blood with blood vessels, muscles, skin, and so on, have some ability for restoration. Therefore, actual revival of not only aging tissue of the human body's membrane, but the entire human body enclosed within, with all internal organs, might be expected. We assess several aging theories within the context of our model and provide suggestions on how to activate the body's own anti-aging mechanisms and increase longevity. This paper presents some analogies and some distinctions that exist between the living dissipative structure matter and inanimate matter, discusses the aging process and proposes certain aging reversal solutions.     

Unusual responses of nocturnal pineal melatonin synthesis and secretion to swimming: Attempts to define mechanisms

Abstract: The effect of swimming at night on rat pineal melatonin synthesis was compared with that of light exposure at night. Rats were forced to swim at 0030 hr (lights out at 2000 hr) and sacrificed by decapitation 15 and 30 min later, immediately after swimming. Other groups of animals were exposed to white light (650μW/cm²) for 15 and 30 min at same time. Swimming caused a rapid and highly significant drop in the melatonin content in the pineal gland; however, the activity of N-acetyltransferase (NAT), the supposed rate limiting enzyme in the melatonin production, was not changed. Despite the drop in pineal melatonin levels, serum concentrations of the indole remained elevated in the rats that swam. In contrast, melatonin levels in the pineal and serum of light exposed rats fell precipitously, accompanied by a significant suppression of NAT activity. Since we anticipated that the strenuous exercise associated with swimming may induce release of artrial natriuretic peptide (ANP) from the heart, which in turn could cause the release of pineal melatonin, in a second study we injected physiological saline intravenously to stretch the cardiac muscle and release ANP. Three milliliters of normal saline was injected during the day into the jugular vein of anesthetized rats that were pretreated with isoproterenol to stimulate pineal melatonin production. Animals were killed 15 min after the saline injection, and pineal NAT activity and pineal melatonin levels were measured. The saline injections caused no alteration in the elevated levels of either NAT or melatonin. These data suggest that the disparity in pineal NAT activity (which was high) and pineal melatonin (which was low), in animals swum at night, may not be caused by ANP which is released during strenuous exercise such as swimming.     

Melatonin and photoperiodic time measurement: Seasonal breeding in the sheep

Abstract: Well-established circadian physiology supports the view that photoperiodic time measurement utilizes the coincidence between the presence of light and a photosensitive phase of a 'biological clock' to alter reproductive status—the so-called external coincidence model of seasonal breeding. In this review, we examine the mechanism whereby photoperiod interacts with presumed suprachiasmatic nuclei activity to allow endogenous melatonin to normally synchronize reproductive activity to the optimal time of year. The Romney Marsh sheep is particularly explored as an experimental model. It is suggested that the on/off activity of seasonal reproduction may be a robust mechanism able to be predictably manipulated by the judicious use of the light/dark cycle and exogenous melatonin, but firmly based on circadian principles.     

MUTATION FREQUENCY IN NURSES AND PHARMACISTS WORKING WITH CYTOTOXIC DRUGS

Individuals occupationally exposed to cytotoxic drugs may be at risk owing to the effects of these agents on DNA. As an index of DNA damage, in vivo mutations were measured in lymphocytes from 24 oncology nurses or pharmacists and 24 matched controls. Mutation frequency was significantly increased in exposed individuals and appeared to be related to duration of exposure. However, the overall magnitude of the increase was small and its biological significance remains to be determined.     

A single injection of adrenergic agonists enhances pineal melatonin production in Turkish hamsters

Abstract: The purpose of this study was to determine whether the pineal gland of Turkish hamsters (Mesocricetus brandti) responds to adrenergic agonists with an increase in melatonin production, and, if it does, whether the sensitivity of the pineal gland to agonists would differ throughout the dark phase. Adult Turkish hamsters weighing 110–210 g received a subcutaneous injection of isoproterenol (ISO, 1 mg/kg B.W.) or norepinephrine (NE, 1 mg/kg B.W.) at different times of night. Animals exposed to LD 16:8 responded to ISO or NE with increased pineal melatonin content only when injected at dawn, when endogenous melatonin is at basal or near-basal levels. When the 8 hr scotophase was entirely replaced with light, the responsiveness to ISO injections at dawn disappeared. In animals exposed to light from 30 min prior to injection to the time of sacrifice, ISO injections increased pineal melatonin content (P < 0.005, three-way ANOVA), which varied, depending on the specific time of injection (effect of time of night, P < 0.05, three-way ANOVA). These results demonstrate that (1) adrenergic agonists enhance the production of pineal melatonin in Turkish hamsters, (2) this stimulatory effect takes place late, but not early in the 8 hr scotophase, and (3) the adrenergic induction of pineal melatonin production in Turkish hamsters requires priming by darkness during the appropriate circadian phase.     

Immune effector mechanisms in malaria: An update focusing on human immunity

The past decade has witnessed dramatic decreases in malaria‐associated mortality and morbidity around the world. This progress has largely been due to intensified malaria control measures, implementation of rapid diagnostics and establishing a network to anticipate and mitigate antimalarial drug resistance. However, the ultimate tool for malaria prevention is the development and implementation of an effective vaccine. To date, malaria vaccine efforts have focused on determining which of the thousands of antigens expressed by Plasmodium falciparum are instrumental targets of protective immunity. The antigenic variation and antigenic polymorphisms arising in parasite genes under immune selection present a daunting challenge for target antigen selection and prioritization, and is a given caveat when interpreting immune recall responses or results from monovalent vaccine trials. Other immune evasion strategies executed by the parasite highlight the myriad of ways in which it can become a recurrent infection. This review provides an update on immune effector mechanisms in malaria and focuses on our improved ability to interrogate the complexity of human immune system, accelerated by recent methodological advances. Appreciating how the human immune landscape influences the effectiveness and longevity of antimalarial immunity will help explain which conditions are necessary for immune effector mechanisms to prevail.     

Secondary aortoduodenal fistula

Aorto-duodenal fistulae （ADF） are the most frequent aorto-enteric fistulae （80%）, presenting with upper gastrointestinal bleeding. We report the first case of a man with a secondary aorto-duodenal fistula presenting with a history of persistent occlusive syndrome. A 59-year old man who underwent an aortic-bi-femoral bypass 5 years ago, presented with dyspepsia and biliary vomiting. Computed tomography scan showed in the third duodenal segment the presence of inflammatory tissue with air bubbles between the duodenum and prosthesis, adherent to the duodenum. The patient was submitted to surgery, during which the prosthesis was detached from the duodenum, the intestine failed to close and a gastro-jejunal anastomosis was performed. The post-operative course was simple, secondary ADF was a complication （0.3%-2%） of aortic surgery. Mechanical erosion of the prosthetic material into the bowel was due to the lack of interposed retroperitoneal tissue or the excessive pulsation of redundantly placed grafts or septic procedures. The third or fourth duodenal segment was most frequently involved. Diagnosis of ADF was difficult. Surgical treatment is always recommended by explorative laparotomy. ADF must be suspected whenever a patient with aortic prosthesis has digestive bleeding or unexplained obstructive syndrome. Rarely the clinical picture of ADF is subtle presenting as an obstructive syndrome and in these cases the principal goal is to effectively relieve the mechanical bowel obstruction.     

Serological survey of HIV and syphilis in pregnant women in Madagascar

Objectives Peripartal transmission of human immunodeficiency virus (HIV) and Treponema pallidum, the causative agent of syphilis, leads to severe consequences for newborns. Preventive measures require awareness of the maternal infection. Although HIV and syphilis testing in Madagascar could be theoretically carried out within the framework of the national pregnancy follow‐up scheme, the required test kits are rarely available at peripheral health centres. In this study, we screened blood samples of pregnant Madagascan women for HIV and syphilis seroprevalence to estimate the demand for systemic screening in pregnancy. Methods Retrospective anonymous serological analysis for HIV and syphilis was performed in plasma samples from 1232 pregnant women that were taken between May and July 2010 in Ambositra, Ifanadiana, Manakara, Mananjary, Moramanga and Tsiroanomandidy (Madagascar) during pregnancy follow‐up. Screening was based on Treponema pallidum haemagglutination tests for syphilis and rapid tests for HIV, with confirmation of positive screening results on line assays. Results Out of 1232 pregnant women, none were seropositive for HIV and 37 (3%) were seropositive for Treponema pallidum. Conclusions Our findings are in line with previous studies that describe considerable syphilis prevalence in the rural Madagascan population. The results suggest a need for screening to prevent peripartal Treponema pallidum transmission, while HIV is still rare. If they are known, Treponema pallidum infections can be easily, safely and inexpensively treated even in pregnancy to reduce the risk of transmission.     

Married to M. tuberculosis: risk of infection and disease in spouses of smear-positive tuberculosis patients

Objectives To quantify the risk of infection and disease in spouses of tuberculosis patients and the extent to which intervention could reduce the risk in this highly exposed group. Methods We compared HIV prevalence, TB prevalence and incidence and tuberculin skin test (TST) results in spouses of TB patients and community controls. HIV‐positive spouses were offered isoniazid preventive therapy (IPT), and TST was repeated at 6, 12 and 24 months. Results We recruited 148 spouses of smear‐positive patients ascertained prospectively and 3% had active TB. We identified 203 spouses of previously diagnosed smear‐positive patients, 11 had already had TB, and the rate of TB was 2.4 per 100 person years(py) over 2 years (95% CI 1.15–5.09). 116 were found alive and recruited. HIV prevalence was 37% and 39% in the prospective and retrospective spouse groups and 17% in controls. TST was ≥10 mm in 80% of HIV negative and in 57% of HIV‐positive spouses ascertained retrospectively; 74% HIV negative and 62% HIV‐positive spouses ascertained prospectively, and 48% HIV negative and 26% HIV‐positive community controls. Of 54 HIV‐positive spouses, 18 completed 6‐month IPT. At 2 year follow‐up, 87% of surviving spouses had TST ≥10 mm and the rate of TB was 1.1 per 100 py (95% CI 0.34–3.29). Conclusions Spouses are a high‐risk group who should be screened for HIV and active TB. TST prevalence was already high by the time the spouses were approached but further infections were seen to occur. Uptake and adherence to IPT was disappointing, lessening the impact of short‐duration therapy.