目的 探讨T细胞免疫球蛋白及粘蛋白结构域3(T cells immunoglobulin domain and mucin domain 3,Tim-3)对单核细胞诱导破骨样细胞生成和骨吸收功能的影响.方法 流式细胞术检测RA患者和体检健康者外周血单核细胞上Tim-3的表达水平;利用人外周血单核细胞体外诱导破骨样细胞;实时定量PCR法检测破骨样细胞形成过程中RANK、CTSK和MMP9 mRNA表达水平;瑞氏染色和肌动蛋白染色观察细胞形态,TRAP染色计数细胞形成数量;采用骨吸收功能试验检测破骨样细胞骨吸收陷窝的数量和面积.结果 RA患者外周血单核细胞上Tim-3表达水平[(77.31±10.66)％]明显高于健康人对照组[(51.72±16.69)％],差异有统计学意义(t=7.593,P<0.01),不同Tim-3水平的单核细胞在诱导为破骨样细胞时,Tim-3高水平组骨吸收陷窝面积[(1.054±0.085)S/mm2]明显低于中间水平组[(1.889±0.053)S/mm2]和低水平组[(2.763±0.066)S/mm2],差异有统计学意义(F=9.318,P<0.05).结论 Tim-3可能对破骨样细胞的骨吸收功能具有负向调控作用.

目的 探讨肠道菌群在3-甲基腺嘌呤(3-MA)改善四氯化碳(CCl4)介导小鼠肝纤维化过程中的作用.方法 15只小鼠随机分为正常对照组、肝纤维化组和3-MA处理组;用CCl4构建肝纤维化模型,3-MA处理组第3周开始额外给予3-MA.8周后处死小鼠并取其血液、肝组织及肠道内容物,分析血清ALT、AST和GGT水平、肝组织病理及肠道菌群情况.结果 3-MA处理组血清ALT和AST明显低于肝纤维化组[(68.6±4.2)U/L vs(111.0±7.8)U/L,(179.0±12.9)U/L vs(253.2±26.7)U/L,P<0.01],且肝组织病变程度减轻.PCoA及NMDS分析将3组小鼠肠道菌群区分.与正常对照组比较,肝纤维化组肠道群落Alpha多样性降低,毛螺菌科丰度显著下降,放线菌门、脱硫弧菌等肠道菌丰度显著升高(P<0.05).与肝纤维化组比较,3-MA处理组肠道群落Alpha多样性增高,毛螺菌科、Blautia菌丰度明显增高,双歧杆菌丰度减低;较正常对照组乳酸杆菌丰度明显增高(P<0.05).结论 3-MA改善了CCl4介导的小鼠肝纤维化,而肠道菌群可能在此过程中起着积极作用.

OBJECTIVE To construct a co-culture system for bone marrow mesenchymal stem cells (BMMSC) and multiple myeloma (MM) cells, and to investigate the effects of co-cultured BMMSC on the migrating and homing of multiple myeloma cells. METHODS The BMMSC from the transgenic mice with green fluorescent protein (GFP) fetal bone were cultured by adherent screening. A co-culture system of BMMSC and MM cell line XG-7 cells was constracted, the proliferation and apoptosis of cells were determined by trypan blue exclusion and Annexin V/PI, respectively, MDC staining was employed to detect the autophagy. The moving direction distribution of molecule in BMMSC and XG-7 cells labeled with PE-CD138 in co-culture process were observed dinamically by confocal microscopy. RESULTS After co-culture with GFP-BMMSC, the resistance of XG-7 cells to apoptosis and autophagy were enhanced; at the same time, their proliferation increased. Apoptosis rates of XG-7 cells directly and indirectly co-cultured with BMMSC were (6.23 ± 0.12)% and (6.97 ± 0.03)% respectively, which were lower than that of XG-7 cells cultured alone (17.90 ± 1.46)% (P < 0. 01). There was low level of autophagy in XG-7 cells co-cultured with BMMSC. XG-7 cells are highly polarized and contained a specialized membrane domain with specific protein and lipid components to contact with BMMSC under confocal microscope. After methyl-β-cyclodextrin treatment, the molecules were normally enriched in the specialized domain. CONCLUSION BMMSC can protect XG-7 cells from apoptosis and autophagy, and obviously promote the proliferation of XG-7 cells, and can influence the migrating and homing of multiple myeloma cells.

Objective To establish our lab′s reference intervals of plasma antithrombin Ⅲ and D‐dimer in dif‐ferent gestational periods of healthy pregnant women .Methods 2 850 pregnant women within different gestational periods and 260 non‐pregnant women were enrolled .The antithrombin Ⅲ (AT‐ Ⅲ ) and D‐dimer(D‐D) were detected by STA‐R evolution automatic coagulation analyzer and original reagents .According to CLSI guideline C28‐A3 ,our lab′s reference intervals were established .Results The reference intervals of plasma antithrombin Ⅲ and D‐dimer in healthy non‐pregnant women were 84 .7% - 123 .3% ,≤ 0 .52 μg/mL ,respectively .The reference intervals of plasma AT‐ Ⅲ at gestational weeks ≤ 13 ,> 13 - 27 ,> 27 were 81 .8% - 115 .8% ,77 .7% - 112 .1% and 68 .1% - 113 .1% , respectively .The reference ranges of D‐dimer concentration were ≤ 0 .75 μg/mL ,≤ 1 .04 μg/mL ,≤ 2 .14 μg /mL ,≤3 .24 μg/mL in group of ≤ 13 weeks ,> 13 - 20 weeks ,> 20 - 27 weeks and > 27 weeks ,respectively .Conclusion The reference values of plasma AT‐ Ⅲ and D‐dimer are determined for each gestational period of pregnancy in our lab ,which may assist clinicians in making accurate clinical evaluation of coagulation ,anticoagulation and fibrinolysis function for pregnancy women .

Objective To investigate the expression and significance of CD56 in the patients with acute myeloid leukemia (AML) .Methods The immunophenotype was detected by multi‐color flow cytometry in 102 AML patients .The expression of CD56 and its correlation with other leukocyte differentiation antigens were analyzed .Results Of 102 AML patients ,CD56 expression was found in 23(22.5% ) cases .The expression of CD56 was positively correlated with lymphoid antigen expression (r=0 .304 ,P<0 .05) ,but had no significant correlation with sex ,age ,and the expressions of CD7 , CD34 ,CD117 and HLA‐DR .The complete remission(CR) rate of the patients with CD56+ AML was significantly lower than that of those with CD56- AML (42.1% vs .74.5% )(χ2 =6 .91 ,P<0 .05) . Logistic multivariate regression analysis showed that CD 56 was an independent factor for CR rate of AML patients(P<0 .05) .Survival analysis showed that disease‐free survival(DFS) of patients with CD56+ AML was shorter than that of those with CD56- AML (χ2 =18 .97 ,P<0 .01 ) .COX regression multivariate analysis demonstrated that CD56 was an independent factor for DFS of AML patients(P<0 .01) .Conclusion The expression of CD56 in AML patients indicates a poor prognosis with a lower CR rate and a shorter DFS .CD56 may be taken as an independent prognostic factor in AML patients .

目的 探讨体外分离培养绿色荧光蛋白(GFP)小鼠骨髓间充质干细胞(BMSCs)的方法,并对GFP-BMSCs进行鉴定.方法 采用贴壁筛选法培养GFP-BMSCs,流式细胞仪检测免疫分型,并用VonKossa试剂盒鉴定细胞的成骨分化能力,油红O染色鉴定细胞的成脂分化能力.结果 分离培养的GFP-BMSCs具有较强的增殖能力和较高的成骨、成脂细胞分化潜能.表型分析显示BMSCs不表达CD34、CD45等造血细胞表面标志,低表达CD106,高表达CD29及CD105等.结论 利用GFP-BMSCs体外培养扩增方法,进而得到一种高表达绿色荧光的BMSCs,成为较为理想、方便、高效的示踪工具,为研究造血干细胞移植的显微解剖关系带来方便.

目的 探讨多发性骨髓瘤继发急性髓系白血病的临床特点及地西他滨的疗效.方法 继发于多发性骨髓瘤(不分泌型)的急性髓系白血病患者1例,女,62岁,经VAD方案化疗后缓解,此后给予苯丙氨酸氮芥维持治疗.3年后患者继发急性髓系白血病,经DA方案治疗后无明显好转,间断予地西他滨治疗2个疗程.结果 患者骨髓原始细胞明显下降,血常规基本恢复正常,此后因为经济原因仅接受支持治疗.结论 多发性骨髓瘤继发急性髓系白血病的发病机制不清,治疗相关和疾病本身因素可能参与其中.该类疾病治疗效果不佳,地西他滨为主的治疗方案可以作为首选.

Purpose To investigate the difference between Her-2 gene amplification and protein expression in breast cancer,and its relationship with hormone-receptor status.Methods With tissue microarray,the differences and correlation between gene amplification and protein expression status of Her-2 in 168 cases of breast invasive ductal cancer was compared,the relationship between gene amplification and protein expression of Her-2 and estrogen receptor(ER),progesterone receptor(PR) and clinicopathologic characteristics was also analyzed.Results Her-2 gene amplification by FISH was observed in 63 of the 168(37.50%) breast cancers.IHC and FISH showed a good coincidence rate in the groups with immunohistochemical scores of 0 /1 +,3 + in breast cancer(Kappa = 0.732,P < 0.01).A low coincidence rate was noted between 2 + and 3 +(Kappa = 0.548,P < 0.01).Her-2 gene amplification was statistically significant correlated with tumor size and lymph node metastasis,while not correlated with the other clinicopathologic features.Additionally,Her-2 gene negatively correlated with the expression of ER(r = 0.161,P < 0.05).Conclusion As an independent prognostic marker,Her-2 gene amplification by FISH has a good coincidence rate in the groups with immunohistochemical scores of 0 /1 +,3 + in breast cancer,while patients with stain scores of 2 + should be testified by FISH.Her-2 gene amplification was statistically significant correlated with tumor size,lymph node metastasis and the expression of ER.

目的 探讨原发性髌骨淋巴瘤的临床特征、影像学表现、诊断、治疗及预后.方法 报道1例以右膝部疼痛为首发症状的病例,并对国内外文献进行复习分析.结果 患者确诊为髌骨弥漫大B细胞淋巴瘤,行放化疗联合治疗后,患者获得完全缓解,随访8个月,患者未见转移复发.结论 原发性髌骨淋巴瘤较为少见,病理类型以弥漫大B细胞淋巴瘤为主,确诊需依靠病理及免疫组织化学,治疗上以放化疗结合的综合治疗效果为佳.

1病历资料患者女,77岁.2010年2月起反复出现胸闷、气急,无发热、咳嗽、盗汗、胸痛、心悸等不适,夜间尚可平卧.既往有“高血压”病史20余年,血压最高达180/110 mmHg(1 mmHg =0.133 kPa),平素服用降压药,血压控制良好.入院体检:神志清楚,消瘦,全身皮肤无黄染,无皮下出血点,球结膜无水肿,颈静脉无怒张,浅表淋巴结未及肿大,胸廓无畸形,左下肺叩诊浊音,呼吸音低,未闻及啰音.

目的:探讨乳腺癌HER-2基因扩增状况与临床病理特征的相关性.方法 采用荧光原位杂交(FISH)方法检测165例乳腺癌石蜡标本的HER-2基因扩增状况,分析基因扩增状况与临床病理特征之间的关系.结果 全组HER-2基因扩增阳性52例(31.5％).不同年龄、肿瘤大小、临床分期及病理类型组间HER-2基因扩增阳性率差异无统计学意义(p＞0.05).HER-2基因扩增阳性率在腋淋巴结转移组明显高于无淋巴结转移组(P＜0.05),p53阳性组明显高于阴性组(P＜0.05),ER、PR阳性组明显低于ER、PR阴性组(均P＜0.01).结论 HER-2基因扩增状况与年龄、肿瘤大小、临床分期及病理学类型无关相性,与腋淋巴结转移和p53基因表达成正相关,与ER、PR表达成负相关.HER-2基因可作为判断乳腺癌预后及拟订治疗方案的良好指标.

<正>原发肺非霍奇金淋巴瘤(PPNHL)是起源于肺内淋巴组织的恶性淋巴瘤,本病较罕见,仅占结外淋巴瘤的3.6%,其中69%~78%起源于支气管黏膜相关的淋巴组织,即原发肺黏膜相关淋巴组织淋巴瘤(pri mary pul monary mucosa-associated

Objective To investigate the correlation of HER-2 gene amplification reported by fluorescence in situ hybridization(FISH) and HER-2 protein expression detected by immunohis-tochemistry(IHC) in breast cancer.Methods HER-2 gene amplification and HER-2 protein expression in paraffin-embedded tissues specimens from 134 patients with breast cancer were examined with FISH and IHC,respectively,and the correlation between the results reported by two techniques were analyzed.Results All of 45 cases with negative HER-2 protein expression were negative in HER-2 gene amplification.Of 33 patients with HER-2 protein expression(+),four cases showed positive HER-2 gene amplification.Of 44 patients with HER-2 protein expression(),28 cases showed positive HER-2 gene amplification.Of 12 patients with HER-2 protein expression(),11 cases showed positive HER-2 gene amplification.The results reported by two methods were significantly correlated with a total accordance rate of 84.3%(P0.01).Conclusion IHC can be used as a method for screening HER-2 expression status in breast cancer.A high accordance exists between the result reported by IHC and that by FISH when HER-2 protein expression is(-) or(),whereas the patients with HER-2 protein expression(+) or () by IHC should undergo FISH test further in order to conferm HER-2 gene expression status.

多发性骨髓瘤(MM)其特点是恶性浆细胞在骨髓微环境中不规则积聚引起的不同程度贫血、骨破坏、肾功能不全、高钙血症和感染[1].MM发病机制尚不明确且目前仍被认为不可治愈,因此建立MM模型具有重大意义.Yaccoby等[2]曾建立起较经典的SCID-hu MM模型,但是该模型却忽略了MM是在有免疫力的人体发生的.我们试用尚有残余免疫力的BALB/c裸鼠建立MM模型并研究其特性.

t(8;21)(q22;q22)是急性髓系白血病中最常见的一种非随机染色体易位,约40%～50%的AML-M2(FAB分型)患者存在此种染色体易位[1].t(8;21)(q22;q22)形成AML1ETO融合基因,虽然AML1-ETO融合蛋白能够改变基因的表达及造血细胞的增殖,但它并不能直接导致白血病的发生[2].

Objective To compare the curative effect of T-VAD regimen and VAD regimen in the treatment of multiple myeloma.Methods Sixty-four multiple myeloma patients were randomly divided into VAD group,VAD plus thalidomide respectively.The beginning dose of thalidomide was 200mg/d, increased 100mg every week till the intolerant dose or below 400mg/d.Results The overall response rate of 83.3% in the T-VAD group was higher than that of 58.8% in the VAD group(P0.05).Conclusion The overall response rate of T-VAD regimen is higher than that of VAD regimen,it would be valuable to do clinical research and widespread popularization.

例1,男,58岁.因四肢皮肤大片瘀斑伴关节肿痛4d于2000年7月31日入院.患者既往体健,4d前针灸、拔火罐后出现四肢皮肤瘀斑、膝及踝节肿痛并活动受限,进行性加重.入院后体检:贫血貌,四肢皮肤尤其是关节处大片瘀斑,右膝关节、踝关节肿胀压痛.

目的 探讨急性白血病(AL)患者治疗前后血清白细胞介素(IL)3、IL-6、IL-8水平的变化及意义.方法 采用酶联免疫法(ELISA),对94例初诊AL患者治疗前后血清IL-3、IL-6、IL-8水平进行检测.结果 急性淋巴细胞白血病(ALL)与急性髓细胞白血病(AML)初诊患者血清IL-3水平均明显低于正常对照组(P＜0.01),而血清IL-6、IL-8水平均明显高于正常对照组(P＜0.01);经治疗ALL完全缓解(CR)患者血清IL-3、IL-6、IL-8水平恢复正常,而AML-CR患者血清IL-3水平较初诊时明显升高(P＜0.01),但仍明显低于正常对照(P＜0.01),IL-6、IL-8水平恢复正常;ALL未缓解(NR)及AML-NR组与初诊组相比无性差异(P＞0.05).AML初诊患者血清IL-3水平与ALL-初诊患者无显著性差异,IL-6水平明显低于ALL患者(P＜0.05),而IL-8水平明显高于ALL患者(P＜0.01).AML各亚型之间血清IL-3水平无显著性差异;M1及M5患者血清IL-6水平明显高于其他亚型(P＜0.01);M4及M5患者血清IL-8水平明显高于其他亚型(P＜0.01).结论 IL-3、IL-6、IL-8与AL发生发展有关,其水平与AL类型有关,观察IL-3、IL-6、IL-8水平的变化可作为判断AL患者病情及疗效的辅助指标.

目的分析引起全血细胞减少的病因,了解全血细胞减少症在各种疾病中的分布,以提高诊断的准确率。方法对371例初诊时全血细胞减少患者的临床资料进行回顾性分析。结果371例患者中,引起全血细胞减少的原因主要为造血系统疾病,共274例(73.9%),其中以再生障碍性贫血最多见,共92例(24.8%);余依次为骨髓增生异常综合征57例(15.4%),其中MDS-RA33例、MDS-RAS3例、MDS-RAEB16例、MDS-RAEB-T5例;急性白血病39例(10.5%),其中急性早幼粒细胞白血病20例;巨幼细胞性贫血22例(5.92%);溶血性贫血16例(4.31%),其中阵发性睡眠性血红蛋白尿9例,自身免疫性溶血性贫血7例;多发性骨髓瘤13例(3.50%);脾功能亢进8例(2.16%);特发性血小板减少性紫癜7例(1.89%);缺铁性贫血6例(1.62%);恶性组织细胞病5例(1.35%);噬血细胞综合征4例(1.08%);淋巴瘤3例(0.81%);原发性骨髓纤维化2例(0.54%)。非造血系统疾病引起的全血细胞减少共97例(26.1%),依次为急、慢性肝病37例(9.97%),感染性疾病22例(5.93%),恶性肿瘤15例(4.04%),其中癌细胞骨髓浸润8例,结缔组织病9例(2.43%),肾脏疾病4例(1.08%),甲状腺机能亢进3例(0.81%),未明原因者7例(1.89%)。结论引起全血细胞减少的原因主要为造血系统疾病,对于全血细胞减少的患者必须进行全面详细的检查,才能提高诊断的准确率。