免疫相关性全血细胞减少症的研究进展

免疫相关性全血细胞减少(IRP)或血细胞减少(IRH)症是从骨髓衰竭性疾病中分离出的一种病理机制相对清晰的独立疾病体系,是由于T淋巴细胞调控失衡导致B淋巴细胞数量、亚群、功能异常,进而产生抗骨髓未成熟造血细胞自身抗体并破坏或抑制之,最终导致外周血细胞减少的一类疾病。诊断IRP/IRH关键依据是测得骨髓造血细胞膜结合自身抗体,或经足量肾上腺皮质激素或(和)大剂量静脉丙种球蛋白治疗有效,以及除外检查。目前主要治疗方法为免疫抑制、促造血及对症支持等综合治疗。     

免疫相关性血细胞减少患者血清细胞因子变化与骨髓免疫的相关性

目的检测89例免疫相关性血细胞减少（IRH／IRP）患者外周血细胞因子的变化状况,观察骨髓细胞活化状态,探讨IRH／IRP的免疫相关机制。方法ELISA方法检测IRH／IRP患者外周血清IL-2、IL-4、IL-6和IFN-γ的含量,分析患者组和正常对照组间细胞因子含量的差异,并对患者组Thl类细胞因子与Th2类细胞因子变化的相关性和升高率以及骨髓细胞免疫活化状况与细胞因子变化的相关性进行分析。结果89例患者血清IL-4、IL-6和IFN-γ呈现不同程度明显升高（P〈0．01）,其中IFN-γ显著高于正常对照组,分别为（38．75±3．67）pg／ml和（8．69±1．68）pg／ml。患者Th1活化相关细胞因子的升高率与Th2活化相关细胞因子的升高率相比具有统计学差异。结论IFN-γ的变化与IRH／IRP者细胞免疫活化密切相关,IL-4和IL-6可能参与与IRH／IRP体液免疫的活化,感染等可加重多重免疫活化,细胞因子含量和骨髓细胞免疫状况的监测有助于IRH／IRP的诊断并有利于评价患者的免疫机能状态。     

免疫相关性血细胞减少症27例临床分析

目的观察27例免疫相关性血细胞减少症（IRH）或全血细胞减少症（IRP）患者的临床特点,以提高对该病的认识。方法分析27例IRH/IRP患者临床表现、血常规、骨髓象、骨髓单个核细胞抗人球蛋白（BMMNC-Coombs）试验、溶血试验、抗核抗体ANA和ENA抗体谱、T淋巴细胞亚群、染色体核型等以及对肾上腺糖皮质激素、环孢霉素A等免疫抑制剂的治疗反应。结果该组病例临床主要表现为贫血、出血、感染;血象呈三系、两系或一系减少,网织红细胞百分比不低;骨髓增生以活跃或明显活跃为主（81.5%）,红系比例多增高;BMMNC-Coombs试验阳性率为70.4%;6例（22.2%）合并系统性红斑狼疮（SLE）等自身免疫性疾病;对肾上腺糖皮质激素、环孢霉素A等治疗有良好反应。结论 IRH/IRP是由于T淋巴细胞调控异常致B淋巴细胞产生针对骨髓未成熟造血细胞的自身抗体而使血细胞减少,BMMNC-Coombs试验对IRH/IRP的诊断具有较高的灵敏性和特异性,多数患者对肾上腺糖皮质激素等免疫抑制剂反应良好。     

免疫相关性血细胞减少症

<正>机体的细胞和体液免疫参与造血调控,维持血细胞在正常水平。当免疫功能失调,特别是对造血系统的负调控亢进,抑制血细胞生成或破坏不同阶段的血细胞,可表现为各种类型的广义的免疫相关性血细胞少症,如自身免疫性溶血性贫血(AIHA)、免疫性粒细胞减少症、免疫性血小板减少性紫癜(ITP)、血小板生成素(TPO)自身抗体导致的低巨核细胞性血小板减少、抗vWF裂解酶(vWFcp)自身抗体导致的血栓性血小板减少性紫癜(TTP)、伊文氏综合征、红细胞生成素(Epo)自身抗体导致的纯红细胞再生障碍(PRCA)、ABO血型不合输血性溶血性贫血等。近年发现一类抗骨髓造血细胞(未成熟血细胞)自身抗体导致的一系、两系或三系血细胞减少,暂称之为狭义的“免疫相关性血细胞减少(IRH)”或“免疫相关性全血细胞减少症(IRP)”。该症以往常被误诊为“不典型再障”、“增生性再障”、“再障早期”,20世纪80年代以后,多归之于骨髓增生异常综合征(MDS)。本文仅介绍IRH/IRP。     

免疫相关性血细胞减少症血液及细胞形态学诊断的初步探讨

目的探讨免疫相关性血细胞减少症的血液学及骨髓细胞形态学特征，提高诊断的准确性。方法对25例IRP／IRH患者的血常规、网织红细胞计数、骨髓细胞形态学、骨髓病理学及骨髓细胞染色体检查的形态学资料进行回顾分析。结果血象：全血细胞减少60％（15／25），两系细胞减少40％（10／25）。贫血占72％（18／25），网织红细胞平均值为2．15％。血小板减少占96％（24／25），白细胞减少占88％（22／25），中性粒细胞百分比或绝对值正常或增高者占92％（23／25），见到晚幼红细胞者占28％（7／25）。骨髓分析：骨髓有核细胞增生程度为活跃、明显活跃，占88％（22／25），有核细胞增生减低者占12％（3／25）。粒系比值减低占64％（16／25），比值正常和升高占28％（7／25）。红系增生活跃，明显活跃者100％，见幼红细胞造血岛者占44％（11／25）。粒，红两系均有病态改变。巨核系：巨核细胞数目正常，增多者占80％（20／25）；数目减少者占加％（5／25），巨核细胞成熟障碍占96％（24／25）。骨髓活检：红系或（和）粒系，巨核系增生表现占60％（15／25），减低表现占40％（10／25）。骨髓细胞染色体检查：25例均核型正常。结论IRP／IRH患者外周血象表现有共性，骨髓细胞形态学改变缺乏特异性，骨髓细胞形态学、骨髓病理学活检及骨髓细胞染色体检查三种方法各有所长，可以互补，联合应用有助于临床诊断和鉴别诊断。     

免疫相关性血细胞减少症--一种新认知的疾病(下)

5 诊断及鉴别诊断 5.1 诊断标准 5.1.1 拟诊标准血象三系或两系、一系血细胞减少,但网织红细胞或(和)中性粒细胞百分比不低;骨髓红系或(和)粒系百分比不低,或巨核细胞不少,易见红系造血岛或嗜血现象;除外了其他原、继发血细胞减少症.符合以上条件者可拟诊IRP(三系血细胞少)或IRH(一系或两系血细胞少).     

免疫相关性血细胞减少症23例临床分析

目的探讨免疫相关性血细胞减少症（IRP/IRH）的发病机制及预后。方法对我院收治的23例患者病例进行分析。结果 23例中并发甲状腺功能亢进、干燥综合征各2例,桥本甲状腺炎、系统性红斑狼疮、糖尿病各1例;网织红细胞正常或增高20例,减低3例;乳酸脱氢酶活性增高14例;外周血单核细胞百分比增高19例,形态异常5例;骨髓单核细胞百分比增高9例,形态异常8例;骨髓增生活跃22例,增生减低1例;白细胞碱性磷酸酶阳性率减低16例,增高3例;单用或联合糖皮质激素、环孢素A及丙种球蛋白为该疾病的主要治疗方法,总有效率为95.6%。结论单核细胞百分比增高及形态异常是IRP/IRH特征的新发现。     

骨髓细胞形态学检查在全血细胞减少症中的临床应用

目的对骨髓细胞形态学检查在全血细胞减少症中的应用价值进行探讨。方法取50例全血细胞减少症患者的骨髓涂片标本参与此次研究,患者均于2017年1月至2018年1月入院进行细胞学检查。对患者的骨髓细胞形态学检查结果予以观察,并结合患者临床表现判断患者的全血细胞减少症的病因。结果 50例患者检查显示,有35例患者病因为造血系统疾病,占70%。包括再生障碍性贫血、骨髓增生异常综合征、急性白血病、溶血性贫血、多发性骨髓瘤、缺铁性贫血。有13例患者病因为非造血系统疾病,占26%。包括急慢性肝病、恶性肿瘤骨转移、感染性疾病。有2例患者无法确诊,确诊率为96%。结论对全血细胞减少症患者实施骨髓细胞形态学诊断,可有效明确患者的具体病因,有利于医师对疾病的诊治。     

嗜血细胞综合征骨髓细胞形态学的诊断及鉴别诊断（附病例40例）

嗜血细胞综合征(henophagocytie syndrome,HPS)是一类骨髓细胞形态学检查常见的骨髓像表现,患者临床表现多样.临床体征多为发热、肝脾或(和)淋巴结肿大、全血细胞减少、肝功能异常和凝血障碍[1].本病包括遗传性和继发性两大类,临床上以继发性更为常见.该病本质上属于良性疾病,但由于其病因多样,临床表现和病理改变与一些恶性疾病如恶性组织细胞病(malignant histiocytosis,MH)相似,骨髓细胞形态学检查对嗜血细胞综合征具有重要的诊断和鉴别诊断价值.我院自2003年1月至2008年1月共检出骨髓HPS40例,回顾分析报告如下.     

全血细胞减少症130例病因分析

全血细胞减少症是多种疾病造成的一种全血减少综合征。血常规检测血红蛋白〈l00g／L,血小板〈100×10^9／L,白细胞〈4．0×10^9／L至少有2次,这种血象的改变常见于造血系统疾病,也见于非造血系统疾病,病种繁多,病因复杂,骨髓细胞学检查对全血细胞减少症患者的病因诊断有重要的价值,结合临床资料可提高临床诊断的准确率。现对我院2009年12月至2010年11月收治的130例全血细胞减少症患者进行回顾性总结和病因学分析,报道如下。     

UniPron is A Fully Effective Non-hormonal Reversible Contraceptive in Baboon Model （Papio Anubis）

Objective To determine the safety and efficacy of UniPron as a reversible contraceptive.
Methods Vaginal swabs were obtained before and after UniPron administration, cultured onto appropriate culture media and bacteria identification was done based on type of media used, Gram stain reactions, colony morphology and biochemical tests. Vaginal biopsy tissues were processed using paraffin wax method, stained with hematoxylin and eosin and examined under light microscopy to determine the effect of the product on vaginal tissues. The effect of UniPron on sperm was examined by mixing the product with electroejaculated spermatozoa in vitro at different concentrations. For efficacy studies, male baboons of proven fertility were mated with UniPron treated or untreated females of proven fertility during the fertile stages.
Results All the five females （100%） that were treated with UniPron did not conceive and they regained total fertility when the treatment was stopped while all the controls conceived. At a concentration of 40%, UniPron completely immobilized spermatozoa in an in-vitro system. UniPron mechanism of action was by lowering the vaginal pH and on application in baboon, the pH was lowered for at least 3 h after which it went back to normal.
Conclusions As we plan for a study to test UniPron as a microbicide to prevent STIs including HIV, our current study has established that this novel product is effective in contraception and harmless to vaginal tissues and vaginal microbial flora in a baboon model （Papio anubis）.     

Evaluation of diuretic and laxative activity of hydro-alcoholic extract of Desmostachya bipinnata （L,） Stapf in rats

OBJECTIVE： In continuation to the growing evidence for therapeutical potential of Desmostachya bipinnata （Linn） Stapf, the current pharmacological study was carried out to evaluate the diuretic and laxative activity of its hydro-alcoholic extract in rats. METHODS： The hydro-alcoholic extract of D. bipinnata whole plant was prepared by using Sox- hlet extractor and subjected to analysis by standard preliminary phytochemical tests. Evaluation of both diuretic and laxative activity was carried out using standard methods as reported earlier. Frusemide （20 mg/kg） was served as positive control for diuretic activity and sennosides （10 mg/kg） served as negative control for laxative activity. RESULTS： The hydro-alcoholic extract showed significant diuretic activity and was found to be the most potent in increasing the urinary output at 500 mg/kg when the effect was compared with that of the standard frusemide （P〈0.01）. Moreover, this extract was found to be most effective in increasing urinary electrolyte concentration （Na＋, K＋, and Cl-） at both doses tested. Whereas the results for laxative activity showed minimal increase of feces output at the dose of 500 mg/kg and the increase was negligible when compared with that of the standard drug sennosides. CONCLUSION： Altogether, the above significant findings validate and support its folkloric diuretic use and lend pharmacological credence to the ethno-medical use of this plant in traditional system of medicine, which demands further studies to investigate its active constituents, as well as its use and safety.     

Effect of the disruption of three cytoskeleton components on chondrocyte metabolism in rabbit knee cartilage

Background Chondrocytes＇ phenotype and biosynthesis of matrix are dependent on having an intact cytoskeletal structure.Microfilaments,microtubules,and intermediate filaments are three important components of the cytoskeletal structure of chondrocytes.The aims of this study were to determine and compare the effects of the disruption of these three cytoskeletal elements on the apoptosis and matrix synthesis by rabbit knee chondrocytes in vitro.Methods Chondrocytes were isolated from full-thickness knee cartilage of two-month-old rabbits using enzymatic methods （n=24）.The isolated cells were stabilized for three days and then exposed to low,medium,and high doses of chemical agents that disrupt the three principal cytoskeletal elements of interest：colchicine for microtubules,acrylamide for intermediate filaments,and cytochalasin D for actin microfilaments.A group of control cells were treated with carrier.Early apoptosis was assessed using the Annexin-FITC binding assay by flow cytometry on days 1 and 2 after exposure to the disrupting chemical agents.The components and distribution of the cytoskeleton within the cells were analyzed by laser scanning confocal microscopy （LSCM） with immunofluorescence staining on day 3.The mRNA levels of aggrecan （AGG） and type Ⅱ collagen （Col-2） and their levels in culture medium were analyzed using real-time PCR and enzymelinked immunosorbent serologic assay （ELISA） on days 3,6,and 9.Results In the initial drug-dose-response study,there was no significant difference in the vitality of cells treated with 0.1 μmol/L colchicine,2.5 mmol/L acrylamide,and 10 μg/L cytochalasin D for two days when compared with the control group of cells.The concentrations of colchicine and acrylamide treatment selected above significantly decreased the number of viable cells over the nine-day culture and disrupted significantly more cell nuclei.Real-time PCR and ELISA results showed that the mRNA levels and medium concentrations of AGG and Col-2 were significantly decreased for     

Liuwei Dihuang,a traditional Chinese herbal formula,suppresses chronic inflammation and oxidative stress in obese rats

OBJECTIVE： To investigate the anti-inflammatory, anti-oxidative stress, and adipokine-ameliorating effects of Liuwei Dihuang （LWDH）, a traditional Chinese herbal formula, in obese rats. METHODS： After 2 weeks of acclimation with free access to regular rodent chow and water, obese-prone-caesarean-derived （OP-CD） rats were fed a modified AIN-93G diet containing 60% energy from fat. Treatment was performed twice daily by gavage feeding with 500, 1 500, or 3 500 mg/kg body weight LWDH suspended in water （n=12 rats per group）. Twelve obese-resistant-CD （OR-CD） rats were fed the atherogenic diet and gavaged with water, and served as the normal control. Blood biomarkers of inflammation, oxidative stress and adiponectin were measured post-sacrifice and used to determine the treatment effect of LWDH and assess the suitability of OR/OP-CD rats for studying these parameters. RESULTS： After 9 weeks of treatment, LWDH lowered serum C-reactive protein （CRP） and tumour necrosis factor-α （TNF-α） levels. Serum interleukin-6 （IL-6） levels showed a tendency towards reduction, but were not significantly different from the OP-CD control. Liver superoxide dismutase （SOD） activity was increased in response to all three doses of LWDH, while the levels of reduced （GSH） and oxidized glutathione （GSSG） and thiobarbituric acid reactive substances （TBARS） were unchanged. Serum adiponectin levels were increased in response to oral administration of LWDH at the dose of either 500 or 1 500 mg/kg body weight. In addition, comparisons between OR-CD and OP-CD rats revealed differential, and for some biomarkers, conflicting characteristics of high-fat diet-fed OP-CD rats in reference to obese human subjects in terms of inflammatory and oxidative stress biomarkers and circulating adiponectin levels. CONCLUSION： The results show, for the first time, the anti-inflammatory, anti-oxidative stress and adiponectin-ameliorating effects of LWDH in obese rats. The suitability of the OP-JOP-CD rat model as     

Iloprost inhibits fracture repair in rats

Background Previous studies have shown that prostaglandins （PGs） dramatically stimulate healing processes in bone.However,the effect of prostaglandin l2 （PGI2） on fracture healing remains unclear.To investigate the effect of PGI2,a study on fracture healing process in closed tibia fractures was designed.Methods Thirty-six Sprague-Dawley male rats were randomized into two groups.On the first day,their right tibias were fractured by three-point bending technique.The study group （n=18） received a single injection of 10 μg/kg iloprost for 5 days,while the control group （n=18） received saline solution in the same way.On the 7th,14th and 28th days following the fracture,six rats were sacrificed and their right legs were harvested in each group.The progression of fracture healing was assessed for each specimen by the scores of radiography （by Lane-Sandhu） and histology （by Huo et al）.Results On the 7th day,the radiographic and histologic scores were equal.On the 14th day radiographic total score was 6 and histologic total score was 23 in the iloprost group,whereas radiographic total score was 11 and histologic total score was 33 in the control group.On the 14th day radiographic and histologic scores were significantly decreased in the iloprost group compared to the control group （P 〈0.05）.On the 28th day radiographic total score was 12 and histologic total score was 37 in the iloprost group,whereas radiographic total score was 15 and histologic total score was 40 in the control group.On the 28th day although there was a decrease in radiographic and histologic scores of the iloprost group acording to control group,it was not statistically significant （P 〉0.05）.Conclusion Iloprost delays fracture healing in early stage in rats.     

Effect of Shenfu on the Expression of Bcl-2 and Bax of Intestinal Mucosal Epithelial Cells after Ischemic Reperfusion in Rats

Objective To study the effect of Shenfu on the expression of bax and bcl-2 in small intestinal mucosal epithelial cells after ischemic reperfusion injury and explore the effect of Shenfu on small intestinal mucosal epithelial cells apoptosis. Methods 36 SD rats were randomized into three groups, each consisting of 12 rats： Sham group （S group）, Ischemic reperfusion group （IR group）, Shenfu group （SF group）, Ischemic reperfusion models were made by ligated the superior menseneric artery for 1 hour followed by 2 hrs reperfusion. Histological mucosal damage in each group was graded according to Chiu＇s score. Immunohistochemistry detected the expression of bax and bcl-2, and obtained the optical density （OD） value using a color image pattern analysis system, and then calculated the ratio of bcl-2/bax. TUNEL method measured apoptotic intestinal mucosal epithelial cells, and calculated the apoptotic index. Results There were edema and epithelial impairment in part of villus and the integral was higher in SF group than S group （P〈0.05）, but lower than IR group （P〈0.01）. Compared IR group with S group and SF group, the average OD value of bcl-2 and bax had significant statistic difference （P〈0.01）, and the average OD value of bcl-2 in SF group was higher than in S group （P〈0.05）, meanwhile bcl-2/bax was significantly lower in IR group than in S group and SF group （P〈0.01）, but that was higher in SF group than in S group （P〈0.05）. Apoptotic index in IR group was significantly higher than that in other two groups （P〈0.01）, and that in SF group was higher than in S group （P〈0.05）. Conclusion Shenfu can enhance the expression of bcl-2, decrease the expression of bax, meanwhile increase the ratio of bcl-2/bax, inhibit small intestinal mucosal epithetlial cells apoptosis, and protect small intestine mucosal epithetlial after ischemic reperfusion.     

补肾活血方对PCOS大鼠模型卵巢中PAI-1mRNA表达的影响

目的探讨补肾活血方对大鼠PCOS模型卵巢局部纤溶酶原激活物抑制因子-1（PAI—1mRNA）表达的影响。方法选用未成年24d龄SD雌性大鼠60只,随机分为模型组、克罗米酚组、补肾活血方高剂量组、补肾活血方低剂量组、正常对照组5组。用Bogovich法建立大鼠多囊卵巢病理模型。以克罗米酚为对照。用原位杂交法观察补肾活血方对多囊卵巢大鼠局部PAI—1mRNA的影响。结果模型组卵巢局部PAI—1mRNA存在卵泡膜间质细胞显著增高,用补肾活血方高、低剂量与克罗米酚药后,卵巢局部PAI-1mRNA的表达明显降低．差异具有统计学意义（P〈0．05、P〈0．01）。补肾活血方高剂量组与克罗米酚组比较,差异具有显著统计学意义（P〈0．01）;低荆量组与克罗米酚组比较差异无统计学意义（P〉0．05）,补肾活血方高、低剂量组比较,低剂量组卵泡膜间质细胞上PAI-1的基因表达增高更明显,但二者差异无统计学意义（P〉0．05）。结论PAI-1mRNA可能与多囊卵巢综合征的发病机制有关。以补肾活血立法的补肾活血方能降低多囊卵巢大鼠局部PAI—1mRNA的显著增高表达．降低PAI—1mRNA卵巢局部的作用。提示补肾活血方可能通过PAI—1mRNA途径促进卵巢排卵的机制。     

New Mechanism of Herb-herb Interaction between Ginseng and Trogopterus Based on CYPs in Rat Livers

The study was designed to investigate the potential mechanism of herb-herb interaction between ginseng and Trogopterus （Trg） based on Cytochrome P450 isozymes （CYPs） in rat livers. We estimated the influence on CYP1A2, CYP2E1, and CYP3A1/2 activity caused by ginseng and Trg used in combination. The CYP1A2 and CYP2E1 enzyme activity were induced by ginseng and Trg used in combination. And this induction effect was caused via inducing CYP1A2 and CYP2E1 protein expression which was supposed caused by inducing the gene expression of CYP1A2 and CYP2E1.     

益智健脑颗粒联合针灸对阿尔茨海默病大鼠学习记忆的影响

目的观察益智健脑颗粒联合针灸对阿尔茨海默病（Alzheimers disease,AD）大鼠学习记忆的影响。方法将大鼠随机分为假手术组（A组）、模型组（B组）、针灸组（C组）、益智＋针灸组（D组）各10只,B、C、D 3组分别以海马CA1区注射β淀粉样蛋白25-35（Aβ25-35）造模,A组注射等量的双蒸水,各组分别治疗20 d后行Morris水迷宫试验,观察大鼠学习记忆能力变化。结果B组较A组的平均潜伏期明显延长,差异具有统计学意义（P〈0.05）;与B组比较,C组、D组的平均潜伏期明显缩短,过台次数增多,差异具有统计学意义（P〈0.05,P〈0.01）;与C组比较,D组的潜伏期缩短,过台次数增多,差异具有统计学意义（P〈0.05）。结论益智健脑颗粒联合针灸能够提高Aβ25-35介导的AD模型大鼠的学习记忆能力。     

The Targeted Pro-apoptotic Effect of scFv-mediated Delivery of Caspase-6 on HER2-positive Osteosarcoma Cells

Objective To investigate the pro-apoptotic effect of scFv-mediated delivery of caspase-6 fusion protein on osteosarcoma E 10 cells. Methods The expression of human epidermal growth factor receptor-2 （Her-2） in osteosarcoma cell line E10 was examined by flow cytometry analysis. ScFv-mediated delivery of caspase-6 was generated by sequential fusion of the genes of a signal peptide, a single-chain Her-2 antibody （e23sFv）, a PEA translocation domain （PEA aa253-264）, and an recombinant caspase-6. The pCMV-immunocaspase-6 was produced by cloning above mentioned fusion genes into pCMV plasmid, and then pCMV-immunocaspase-6 plasmid mixed cationic liposome was transfected into E10 cells. The expression of fusion gene on transfected E10 cells was detected by immunocytochemical staining, The pro-apoptotic effects of fusion gene expression on morphology and growth status of E10 cells were observed by trypan blue cell staining, electron microscopy and MTT assay. Results The fusion protein was detected in the cytoplasm of the transfected E10 cells. These cells presented typical characteristics of apoptosis as detected by electron microscopy. MTT assay revealed that the proliferation of transfected E10 cells was suppressed than that of non- or mock-transfected cells （P〈0.01）. Conclusion ScFv-mediated delivery of caspase-6 fusion protein can induce apoptosis of Her-2 positive osteosarcoma E10 cells, suggesting the potential of this strategy for the treatment of human high-grade osteosarcoma.