自体外周血造血干细胞移植治疗难治性系统性红斑狼疮

目的:探讨自体外周血造血干细胞移植(APBSCT)治疗难治性系统性红斑狼疮(SLE)的临床疗效。方法:用APBSCT治疗4例难治性SLE。干细胞动员应用环磷酰胺(CTX)4g/m2,分两天应用和粒细胞集落刺激因子(G-CSF)5~10μg/kg·d-1;预处理方案包括CTX(50mg/kg·d-1,-6、-5、-4、-3d),抗胸腺细胞球蛋白(ATG15~20mg/kg·d-1,-2、-1、 1、 2d)。结果:4例患者均获得造血重建,中性粒细胞>0.5×109/L,血小板>20×109/L的中位数时间分别是9d,10d;SLE的临床表现明显减轻,尿蛋白减少或消失,自身抗体转阴或滴度减低,泼尼松用量<10mg/d;无移植相关死亡。结论:APBSCT治疗难治性SLE近期疗效显著,造血重建恢复迅速,安全有效,远期疗效尚需进一步观察。     

自体外周血造血干细胞移植治疗系统性红斑狼疮的临床观察

目的 探讨自体外周血造血干细胞移植(APBSCT)治疗难治性系统性红斑狼疮(SLE)的临床疗效和安全性.方法 10例难治性SLE患者接受APBSCT治疗,应用环磷酰胺(CTX)2～4 g/m2和粒细胞集落刺激因子5～10 μg·kg-1·d-1行外周血造血干细胞动员;预处理包括CTX(50 mg·kg-1·-1,-6～-3 d)和抗胸腺细胞球蛋白(ATG,15～20 mg·kg-1·d-1,-2 d、-1 d、1 d、2 d).患者输注的CD34+细胞＞2×106/kg.评估治疗前后临床表现、SLE疾病活动指数(SLEDAI)和免疫指标的变化.结果 APBSCT后10例SLE患者的临床症状缓解,SLEDAI评分降低,均获得造血重建,中性粒细胞＞0.5×109/L的中位数时间为9.5 d,血小板＞20×109/L中位数时间是11 d;尿蛋白减少或消失,抗核抗体滴度减低或转阴,补体水平升高;移植相关的并发症有:2例败血症,2例巨细胞病毒感染,1例出现肾毒性,3例急性左心衰竭,3例心律失常,无移植相关死亡.结论 APBSCT能够改善SLE患者的疾病活动和免疫学指标,是一种有效的治疗难治性SLE的方法,但远期疗效需进一步观察.     

自体骨髓干细胞移植治疗系统性红斑狼疮的临床研究

目的：探讨自体骨髓干细胞移植(ABMSCT)对系统性红斑狼疮(SLE)的临床疗效。方法：3例患者采集自体骨髓移植CD34^ 造血干细胞，预处理用环磷酰胺(CTX)60mg／kg连续2d静脉滴注，马法兰140mg／m^2分次口服，抗CD3、CD8单抗各5mg／d连续5d静脉滴注，用粒细胞集落刺激因子(G-CSF)协助造血及免疫重建，用间接免疫荧光法检测抗核抗体、放免法测抗DNA抗体、流式细胞仪测淋巴细胞亚群，观察ABMSCT前后临床表现和免疫学指标的变化。结果：ABMSCT后患者的临床症状明显缓解，异常免疫学指标恢复正常，抗体全部转阴。结论：ABMSCT对SLE有较好的近期疗效，远期疗效还需长期随访。     

自体外周血干细胞移植治疗难治性系统性红斑狼疮的临床研究

目的探讨自体外周血干细胞移植(APBSCT)治疗难治性系统性红斑狼疮(SLE)的临床疗效。方法对难治性SLE患者进行APBSCT治疗。采用环磷酰胺(CTX) 粒细胞集落刺激因子(G-CSF)方案动员,CS-3000血细胞分离机采集外周血干细胞并保存于-80℃冰箱;用CTX50mg/(kg.d)×3～4d方案预处理后,解冻后回输冻存的干细胞的治疗方法。观察APBSCT前后临床表现及免疫学指标的变化。结果动员后获得单个核细胞数(MNC)(3.383～3.704)×108/kg;CD34 细胞数(4.4～11.12)×106/kg。3例均获得造血重建,中性粒细胞>0.5×109/L、血小板>20×109/L的中位数时间分别是8.3d、10d。移植后患者临床症状消失,1例并肾功能不全、难治性高血压和重度贫血的患者移植后恢复正常。3例患者自身抗体转阴或滴度减低,尿蛋白定性消失,SLE疾病活动指数(SLE-DAI)由移植前的平均15分下降为移植后6个月的平均4分。结论APBSCT治疗难治性SLE安全有效,近期疗效好,远期疗效有待进一步观察。     

免疫清除性化疗结合自体外周血造血干细胞移植治疗重症系统性红斑狼疮

目的 观察免疫清除性化疗结合自体外周血造血干细胞移植（移植）治疗重症系统性红斑狼疮（SLE）的疗效及安全性。方法 重症SLE4例，分别合并狼疮性肾炎，狼疮脑，狼疮心或股骨头坏死，干细胞的动员采用环磷酰胺加重组人粒细胞集落因子（G－CSF）；预处理为回输前3天每天应用环磷酰胺50mg/kg 回输后3天每天应用抗胸腺细胞球蛋白（ATG）5mg/kg。观察移植前后临床症状，体征，狼疮相关抗体等指标的改变，并动态观察移植后免疫功能的重建。结果 移植后患者的临床症状完全缓解，狼疮相关抗体全部转阴，移植后患者的免疫功能均明显降低，约半年后恢复正常，但不伴有临床症状的复发。结论 免疫清除性化疗结合自体外周血造血干细胞移对难治性SLE有明显的疗效，尤其适用对各种药物治疗无效者，治疗是安全的，远期疗效还需长期随访。     

外周造血干细胞移植治疗自身免疫性疾病疗效随访

目的:研究大剂量环磷酰胺(CTX)免疫清除治疗后用外周血干细胞救助治疗自身免疫性疾病(AD)的安全性及疗效。方法:AD18例,其中严重系统性红斑狼疮(SLE)16例,多发性硬化与晚期类风湿关节炎各1例,用大剂量CTX免疫清除治疗(CTX50mg/kg·d-1×3d)+自身造血干细胞输注救助。结果:18例AD都成功进行了自体外周血干细胞移植(APBSCT)的全过程,造血功能及免疫功能迅速恢复,未出现严重的毒性反应及并发症。16例SLE完全缓解,相关抗体全部阴转,长期随访(最长48个月),2例SLE复发,1例多发性硬化在治疗后9个月复发。结论:大剂量CTX免疫清除治疗后用造血干细胞救助治疗AD,造血功能恢复较快,是安全有效的,持续缓解时间长,复发率低(16%),但还需进一步扩大病例,长期随访。     

外周血干细胞移植治疗难治性系统性红斑狼疮

目的 观察自体外周血干细胞移植（APBSCT）治疗难治性系统性红斑狼疮（RSLE）的疗效及安全性。方法 RSLE3例，先给予血浆置换，环磷酰胺（CTX）和大剂量丙种球蛋白，然后用环磷酰胺和粒细胞集落刺激因子（G-CSF）动员造血干细胞（HSC），多功能加强型血细胞分离机采集外周血干细胞，-86℃超低温冻存，环磷酰胺预处理，经静脉回输干细胞，最后应用抗胸腺淋巴细胞球蛋白（ATG）行体内去T淋巴细胞。观察移植前后临床症状，体征和免疫学指标的变化。结果 APBSCT后患者的临床症状完全缓解，异常免疫学指标基本恢复正常，抗体全部转阴。结论 APBSCT对难治性系统性红斑狼疮有明显的疗效。尤其适用对各种药物治疗无效的患者，远期疗效还需长期随访。     

G-CSF对造血干细胞的动员机制及对健康供者的影响

粒细胞集落刺激因子（G－CSF）通过影响因干细胞粘附分子的表达及其功能，以及影响骨髋微环境，从而促使骨髓造血干细胞向外周血释放，使外周血造血干细胞移植得以实施。G－CSF可引起供者骨痛，疲乏，头痛、恶心，呕吐，发热等不良反应及免疫功能的暂时性改变，绝大多数供者可耐受上述不良反应，但G－CSF对供者的远期影响尚待进一步研究和观察，本文就上述内容作一综述。     

大剂量化疗并自体外周血干细胞移植治疗重症原发性干燥综合征

目的探讨大剂量化疗（HDC）并自体外周血干细胞移植（PBSCT）对于常规治疗反应不佳的重症原发性干燥综合征（pSS）的可行性、疗效及安全性。方法选择1999年起3例长期激素和免疫抑制剂治疗病情不能缓解的重症pSS患者作为治疗对象，给予大剂量免疫抑制剂治疗及自体外周血干细胞移植。自体干细胞动员采用环磷酰胺（CTX）2—3g／m^2和粒细胞集落刺激因子（G—CSF），并行CD34^＋细胞分选。干细胞回输前用CTX200mg／kg、抗胸腺细胞免疫球蛋白（ATG）90mg／kg或CTX200mg／kg、全身照射（TBI）4gy进行预处理。结果1例患者完成2次CTX2g／m^2动员，尚未行干预处理和细胞回输，2例重症pSS顺利完成治疗全过程，3例患者随诊时间分别达48、60和18个月。治疗后3例患者的B淋巴细胞功能亢进均得以抑制，2例患者抗SSB抗体转阴，1例患者重复唇腺活检示灶性淋巴细胞浸润消失：2例患者的肺功能改善，间质性肺炎得以逆转。结论自体外周血干细胞移植可控制重症pSS患者的病情，具有可行性和安全性，疗效较为肯定。     

自体外周血CD34+细胞移植治疗小儿重症自身免疫性疾病的初步观察

目的探讨自体外周血CD34^＋细胞移植治疗小儿重症自身免疫性疾病的疗效问题。方法3例重症自身免疫性疾病患儿接受自体外周血CD34^＋细胞移植。首先采用环磷酰胺（CTX）＋粒细胞集落刺激因子（G—CSF）方案动员外周血干细胞，通过CliniMACS细胞分选仪分选CD34^＋细胞后冻存。然后选用BEAM＋抗胸腺细胞球蛋白（ATG）或CTX＋马法兰（Mel）＋ATG两种方案预处理，0d回输CD34^＋细胞。观察造血重建、免疫恢复以及症状体征变化情况。结果动员获得了足够单个核细胞，纯化后CD34^＋细胞纯度高。粒细胞植入时间分别为9、13、11d，血小板植入时间分别为14、18和13d。3例原发病缓解。随访14～16个月，皮肌炎皮损消失、四肢肌力恢复正常，类风湿关节炎疾病活动评分（DAS28）由7.3分降至2.4分，系统性红斑狼疮DAI评分由16分降至4分。结论自体外周血CD34^＋细胞移植是常规治疗无效的小儿重症自身免疫性疾病的可选择治疗措施之一。     

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.     

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.     

Variabilité des concentrations plasmatiques de l’angiotensinogène et risque d’hypertension artérielle chez le rat transgénique

Aim

Genetic polymorphisms of the human angiotensinogen gene are frequent and may induce up to 30% increase of plasma angiotensinogen concentrations with a blood pressure increase of up to 5 mmHg. Their role for the pathogenesis of human arterial hypertension remains unclear. High plasma angiotensinogen levels could increase the sensitivity to other blood pressure stressors.

Methods

Male transgenic rats with a 9-fold increase of plasma angiotensinogen concentrations and male non-transgenic rats aged 10 weeks were treated or not with NG-Nitro-L-arginine-methyl ester for 3 weeks in their drinking water (n = 3/group). Systolic blood pressure and body weight were measured at baseline and at the end of the study when left ventricular weight and ventricular expression of angiotensin I-converting enzyme and procollagen Iα1 were determined (polymerase chain reaction).

Results

At baseline, transgenic rats had +18 mmHg higher bood pressure and –8% lower body weight compared to non-transgenic rats (P < 0.05) without significant changes for the vehicle groups throughout the study (P > 0.05). NG-Nitro-L-arginine-methyl ester increased blood pressure, left ventricular weight and left ventricular weight indexed for body weight by +41%, +17.6% and +18.6% (P < 0.05) in transgenic and +25%, +5.3% and +6.7% (P > 0.05) in non-transgenic rats compared to untreated animals, respectively. Cardiac gene expression showed no differences between groups (P > 0.05).

Conclusion

Increased plasma angiotensinogen levels may sensitize to additional blood pressure stressors. Our preliminary results point towards an independent role of angiotensinogen in the pathogenesis of human hypertension and associated end-organ damage.     

Morphological and immunocytochemical heterogeneity of cultured pinealocytes from one-week-and two-month-old rats: Planimetric and densitometric investigations

Abstract: In vitro preparations of rat pinealocytes are widely used for biochemical analyses of signal transduction processes. This paper deals with morphological and immunocytochemical features of such preparations. Special attention was paid to the problems of whether pinealocytes represent a heterogeneous cell population and how such heterogeneity may develop during ontogeny. The investigations were performed with cells which were obtained from the pineal organ of one-week-and two-month-old rats, attached to synthetic peptide-coated coverslips or tissue culture chamber slides, and maintained under in vitro conditions overnight. The attached cells were then fixed with paraformaldehyde. These preparations yielded monolayers of spherical cells of different sizes; most cells were isolated, but some of them were aggregated and formed small clusters. On the average, the cells from the one-week-old animals were smaller than the cells from the two-month-old animals. Immunocytochemical demonstration of S-antigen, a pinealocyte-specific marker, showed that the majority of the cells from two-month-old animals were intensely or moderately labelled. Pinealocytes from one-week-old animals were less S-antigen immunoreactive. Only very few cells (less than 1% displayed glial fibrillary acidic protein (GFAP)-immunoreactivity. Planimetric investigations of the cell size and semiquantitative densitometric investigations of the intensity of the S-antigen immunoreaction revealed that (i) pinealocytes kept in vitro form a heterogeneous cell population, and that (ii) this heterogeneity increases during postnatal development from one-week-old to two-month-old animals. Two groups of pinealocytes can be distinguished based on their developmental fate: pinealocytes of one group grow dramatically, but show only a moderate increase in S-antigen immunoreactivity, and pinealocytes of the other group retain their size, but display a distinct increment in S-antigen immunoreacti vitv.     

Effects of pinealectomy and melatonin administration on certain indices of ovarian hyperplasia and/or hypertrophy in rats with both ovaries intact or after unilateral ovariectomy

Abstract: In earlier studies from other laboratories it was shown that melatonin decreased ovarian weight in rats and inhibited compensatory hypertrophy of the remaining ovary after unilateral ovariectomy. This study was designed to examine the influence of melatonin on certain indices of ovarian hyperplasia and/or hypertrophy in adult female rats with both ovaries preserved and with either an intact pineal gland or with the pineal gland removed (pinealectomy, PX) or, finally, in sham-PX animals. Similar studies were conducted on rats after unilateral ovariectomy, referring the examined parameters to the remaining intact ovary. The studies included mitotic activity of granulosa layer cells and corpus luteum cells, ovarian weight, ovarian cross-sectional area, cross-sectional area of the granulosa layer of all the Graafian follicles and the cross-sectional areas of the corpora lutea, visible on the ovarian cross-section. On the basis of results, we conclude that: 1) the effect of PX on the processes of ovarian hyperplasia and hypertrophy may vary; analogously, exogenous melatonin administration may influence ovarian hyperplasia and hypertrophy in different ways; 2) PX and exogenous melatonin may, under certain conditions, exert similar biological effects, even synergistic effects; 3) melatonin inhibits ovarian growth processes, while the effects of PX are variable; 4) the results indicate that in experiments performed on rats, with the use of two control groups, i.e., intact and sham-PX, melatonin effects on these two groups may differ.