目的:探讨自体骨髓干细胞移植对终末期肝病患者的治疗作用.方法:56例终末期肝病患者无菌采集骨髓,负收集法分离纯化骨髓干细胞,经肝固有动脉注入肝脏.术后观察肝脏储备功能和合成功能,同时观察患者症状体征和不良反应.结果:患者骨髓采集量为120 ～ 180al,分离细胞数为5.8×107～9.6×108个,与术前比较,术后1周91.6％的患者食欲改善,87.5％的患者体力明显改善;术后12周吲哚氰绿血浆清除率(ICGK)升高,吲哚氰绿15min滞留率(ICGR15)下降,与术前比较差异有显著性意义(P＜0.05);血清血蛋白(Alb)在术后4周开始升高,12周达到最高,与治疗前比差异有显著性意义(P＜0.05);胆碱酯酶(CHE)、凝血酶原时间(PT)移植术后持续改善,12周时较术前增加最明显,差异有显著性意义(P＜0.05);18例并发腹水者,有11例患者腹水消失,有5例腹水减轻但没有消退.结论:自体骨髓干细胞移植技术治疗终末期肝病,患者肝脏储备功能、合成功能改善明显,术后无不良反应,安全可行.

间充质干细胞（Mesenchymal stem cells,MSCs）是一类具有多向分化潜能的干细胞,具有高度可塑性,在特定的诱导条件下可分化为不同胚层来源的细胞,其生物学特点使其在临床治疗如组织和器官修复等方面具有广阔的应用前景。在一定诱导条件下MSCs可分化为中胚层的骨细胞、软骨细胞、脂肪细胞、心肌细胞,外胚层的神经元、神经

BACKGROUND: Studies have found that cord blood contains a large member of hematopoietic stem cells and abundant mesenchymal stem cells. OBJECTIVE: To establish an isolating, culturing and proliferating method for human umbilical cord blood mesenchymal stem cells (HUMSCs) and then to observe cell biological characteristics in vitro. METHODS: HUMSCs were isolated and purified by density gradient centrifugation and adhering to the culture plastic. After successive subculture and amplification, the morphology was observed under microscope and the growth curve was drawn. Cell cycle was determined by flow cytometry; The expression of CD29, CD44 and CD34 in HUMSCs were detected by immunohistochemistry (SP); HUMSCs were induced to differentiate into adipocytes in DMEM with 20% horse serum and identified by oil red O staining; HUMSCs were induced to differentiate into osteoblasts by 0.1 μmol/L dexamethasone, 10 mmol/L β-glycerol phosphate sodium ascorbate and 50 μmol/L ascorbic acid and identified by alkaline phosphatase staining. RESULTS AND CONCLUSION: High purity HUMSCs were isolated from umbilical cord blood. HUMSCs were spindle cells and their living behavior was quite stable. In vitro subcultured nine generations had no morphological changes and no signs of aging; Passage 3 of HUMSCs could differentiate into adipocytes and osteoblasts; The results showed that density gradient centrifugation, adherent culture and digestion time control can obtain an access to high purity HUMSCs with strong proliferation and passage ability in vitro.

背景:研究发现肝细胞生长因子、成纤维细胞生长因子4和表皮细胞生长因子等可诱导骨髓间充质干细胞向肝细胞分化,关于肝衰竭大鼠血清对骨髓间充质干细胞的诱导分化作用尚未见报道。目的:观察急性肝衰竭大鼠血清对骨髓间充质干细胞的诱导分化作用。方法:采用密度梯度离心法从SD大乳鼠股骨、胫骨分离培养骨髓间充质干细胞,体外扩增培养。采用急性肝衰竭大鼠血清诱导培养骨髓间充质干细胞,以常规培养液诱导为对照,观察细胞形态的变化,诱导培养14d采用免疫细胞化学方法检测甲胎蛋白、白蛋白的表达。结果与结论:急性肝衰竭大鼠血清诱导培养14d,可见骨髓间充质干细胞发生明显的形态学改变,镜下观察细胞变得稍大而扁平,呈类上皮样细胞,甲胎蛋白和白蛋白表达阳性率分别为（54.8±7.64）%和（45.9±9.68）%;对照组细胞未发生形态学改变,无甲胎蛋白和白蛋白的表达。证明了急性肝衰竭大鼠血清对骨髓间充质干细胞的诱导分化作用,可诱导骨髓间充质干细胞表达甲胎蛋白和白蛋白。

目的 观察3,4,5-三羟基苯甲酸(TBA)对荷肝癌H22小鼠生存时间的影响.方法 建立移植性H22肝癌细胞荷瘤小鼠模型,每组10只,TBA高、中、低剂量组TBA溶液0.50、0.25、0.13 g/kg灌胃给药;对照组0.5%羧甲基纤维素钠溶液0.2 ml/10 g灌胃给药;环磷酰胺(CTX)组CTX 20 mg/kg腹腔注射,每天固定时间给药1次,连续10 d,观察各组小鼠生存时间,计算生命延长率.结果 TBA和CTX组小鼠平均生存时间不同程度长于模型组,其中TBA 0.25 g/kg组、CTX组与模型组比较差异显著(P＜0.05),TBA 0.50、0.13 g/kg组与模型组比较差异不显著(P＞0.05).CTX组和TBA组小鼠生命显著延长率分别为29.02%、9.79%、24.13%和6.29%.结论 TBA灌胃给药可以不同程度抑制H22肿瘤的生长,延长荷瘤小鼠生存时间.

Objective To evaluate the diagnostic value on measles virus(MV)from throat swab and urine residue in children measles cases by indirect immunofluorescence assay(IFA).Methods Total of 68 suspected children measles cases were detected for MV antigen from throat swab and urine residue.The positive rate of two kinds of sample were analyzed and some patients were followed-up for short period.Results Total of 62 patients were diagnosed as measles.The positive rates of MV antigen detection in throat swabs and urine residue were 91.9%(57/62)and 96.8%(60/62),respectively,the differences were not statitically significant(χ2 = 1.363,P>0.05).All of 45 patients were diagnosed as typical measles,the MV antigen detection positive rates of throat swabs and urine residue were 95.6%(43/45)and 97.8%(44/45),respectively,the differences were not statitically significant(χ2 = 0.345,P>0.05);There were 17 patients diagnosed as atypical measles,whose MV antigen detection positive rates of throat swabs and urine residue were 82.4%(14/17)and 94.1%(16/17),respectively,the differences were not statitically significant(χ2 = 1.133,P>0.05).Conclusions The detection of MV antigen from throat swab and urine residue by IFA is a rapid and sensitive method.

BACKGROUND:In vitro induced bone marrow mesenchymal stem cells(BMSCs) differentiated into neuron-like cells mainly by polypeptide preparations such as nerve growth factor.Pure chemical inducer is not commonly found.OBJECTIVE:To establish a system for isolation and culture of BMSCs and induce it directional differentiation into neuron-like cells in vitro.METHODS:BMSCs were isolated,purified and identified by density gradient centrifugation,adherent culture and digestion time control.BMSCs were induced into neuron-like cells by β-mercaptoethanol and dimethyl sulfoxide.The morphology change was observed.The differentiation ratio analysis of differentiated neuron-like cells was detected by Nissl staining and immunochemical of neuron specific enolase and neurofilament-200.RESULTS AND CONCLUSION:Separated BMSCs were fibroblast-like cells and they had several nucleoluses.BMSCs could differentiate into neuron-like cells,with the presence of long axons and dendrite-like processes after being induced by β-mercaptoethanol and dimethyl sulfoxide.Both Nissl's body,neurofilament-200 and neuron specific enolase of neurons after induction were positive and the positive rates of neuron specific enolase and neurofilament-200 were(85.6 ±6.7)% and(73.2 ± 5.6)%,respectively.Results demonstrated that density gradient centrifugation,adherent culture and digestion time control can successfully isolate and culture human BMSCs.Human BMSCs can differentiate into neuron-like cells induced by β-mercaptoethanol and dimethyl sulfoxide in vitro.

背景:对于诱导骨髓间充质干细胞成肝细胞样细胞的研究,在大鼠及小鼠及人类中已有相关报道.甲胎蛋白和白蛋白均为肝细胞系较为特异的标志.目的:进一步验证肝细胞生长因子对骨髓间充质干细胞的诱导分化作用.方法:采用密度梯度离心和贴壁培养法相结合分离培养人骨髓间充质干细胞,以含20 μg/L肝细胞生长因子和10 μg/L碱性成纤维细胞生长因子的低糖DMEM培养基诱导培养,对照组取同代细胞,常规培养液培养.诱导培养7,14,21和28d采用免疫组织化学方法检测甲胎蛋白和白蛋白的表达及RT-PCR技术检测白蛋白mRNA表达.结果与结论:诱导组骨髓间充质干细胞呈类上皮样细胞.诱导组在培养7d时,甲胎蛋白表达阳性率为81.5%,随着培养时间延长,甲胎蛋白阳性表达率逐渐减弱,而白蛋白阳性表达率及白蛋白mRNA表达逐渐增强,至培养28d,白蛋白阳性表达率达91.2%,对照组细胞均无白蛋白、白蛋白mRNA及甲胎蛋白表达.结果证实,肝细胞生长因子可诱导骨髓间充质干细胞表达甲胎蛋白和白蛋白.