Expression of aquaporin-1 and aquaporin-3 in lung tissue of rat model with ischemia-reperfusion injury

End-stage lung diseases are common and frequentlyoccurring diseases which are difficult for clinical treatment. In recent years, lung transplantation has become a widely accepted and effective therapeutic option for patients with the end-stage pulmonary diseases. Early pulmonary edema resulting from ischemia-reperfusion injury accounts for the major part of mortality and morbidity after lung transplantation. The water channel proteins in lung injury have been little studied, and their impact on the formation of pulmonary edema remains unclear. In this study, we established a rat lung ischemia-reperfusion model to study its impact on the expressions of water channel proteins in lung tissue and explore a new approach to lung transplantation in pulmonary edema pathogenesis.     

Application of Chinese Traditional Medicine in Kidney Transplantation

Kidney transplantation is one of the important means in saving the life of patients with renal diseases of the terminal stage. However, rejection reaction often results in the loss of the grafted organs' function. Almost half the recipients have been found to suffer from loss of graft function due to rejection reaction within 10 years of post-transplantation. Part of them have bad quality of life     

加强我国儿科领域中对遗传性肾脏病的诊断

More and more hereditary diseases,including hereditary renal diseases in particular,have been diagnosed withthe achievement of the Human Genome Project in the last decade and the development of biotechnology.The in-creased diagnosis of hereditary diseases…     

Small bowel autotransplantation combined with pancreato- duodenectomy for enormous cavernous hemangioma of the small intestine mesentery

Recent advances in transplantation techniques have allowed pancreatoduodenectomy, distal gastrectomy,hemicolectomy and small bowel autotransplantation to be the therapy of choice for enormous cavernous hemangioma of the small intestine mesentery. There have been a few case reports about small bowel autotransplantation combined with pancreatoduodenectomy for enormous mesenteric cavernous hemangioma of small intestine. The present surgical methods for enormous cavernous bemangioma of the small intestine mesentery mainly included tumor excision and/or small bowel resection. However, these therapies are not effective for those patients in whom the angiocavemoma has infiltrated the mesenteric artery or pancreas, and these patients often give up therapy. It is recognized that enormous cavernous hemangioma of the small intestine mesentery is a benign lesion, and patients may have an excellent prognosis after complete resection of the lesion.     

Hepatocyte growth factor gene-modified bone marrow-derived mesenchymal stem cells transplantation promotes angiogenesis in a rat model of hindlimb ischemia

Angiogenic gene therapy and cell-based therapy for peripheral arterial disease (PAD) have been studied intensively currently.This study aimed to investigate whether combining mesenchymal stem cells (MSCs) transplantation with ex vivo human hepatocyte growth factor (HGF) gene transfer was more therapeutically efficient than the MSCs therapy alone in a rat model of hindlimb ischemia.One week after establishing hindlimb ischemia models,Sprague-Dawley (SD) rats were randomized to receive HGF gene-modified MSCs transplantation (HGF-MSC group),untreated MSCs transplantation (MSC group),or PBS injection (PBS group),respectively.Three weeks after injection,angiogenesis was significantly induced by both MSCs and HGF-MSCs transplantation,and capillary density was the highest in the HGF-MSC group.The number of transplanted cell-derived endothelial cells was greater in HGF-MSC group than in MSC group after one week treatment.The expression of angiogenic cytokines such as HGF and VEGF in local ischemic muscles was more abundant in HGF-MSC group than in the other two groups.In vitro,the conditioned media obtained from HGF-MSCs cultures exerted proproliferative and promigratory effects on endothelial cells.It is concluded that HGF gene-modified MSCs transplantation therapy may induce more potent angiogenesis than the MSCs therapy alone.Engraftment of MSCs combined with angiogenic gene delivery may be a promising therapeutic strategy for the treatment of severe PAD.     

Impact of chronic kidney disease on serum tumor markers concentrations

Background Serum tumor markers have always been of clinical importance in the diagnosis,monitoring disease progression and therapy efficacy for patients with malignant diseases.However,elevated serum t...     

Effects of flotation therapy on relaxation and mental state

Flotation therapy is one of the models of Restricted Environmental Stimulation Therapy (REST). It has been studied and used in clinics in western countries for many years. According to the research results, flotation therapy can be used effectively in many ways.1 Some of the research done in China showed that flotation therapy could be helpful in the treatment of hypertension2 as well as cerebral paraplegia. It has also been observed in clinics that flotation therapy can induce deep relaxation, improve emotional states and have beneficial effects on some kinds of neurosis and psychosomatic diseases. But the effect of flotation therapy on the basic psychological and physiological function of ordinary Chinese has not been studied systemically. And there is no objective research result that has demonstrated the benefits of flotation therapy in Chinese clinics.     

Surgical therapy for portal hypertension in patients with cirrhosis in China: present situation and prospects

Surgical therapy for portal hypertension （PHT） in patients with cirrhosis has long been controversial,and-various operative approaches have been used to prevent or manage such lethal complications as gastroesophageal variceal hemorrhage. However, over a century various portazygos devascularization and shunt approaches have been developed or modified, but the therapeutic effects have not been satisfactory till 1963 when Starzl did the first liver transplantation in the world, which provides a new hope to the surgical treatment of PHT.     

Adipose-derived stem cells in stroke treatment: translational possibility and mechanism

Stroke has become one of the secondary worldwide mortal diseases that burdens the family and the society both emotionally and economically. The announcement of China＇s Ministry of Health in 2008 showed that stroke had been the first cause of death in urban and rural residents.1.2 Traditional therapies such as thrombolysis, anti-platelets, and rehabilitation showed limited effects. Replacement therapy with stem cells promises to be a good solution due to its possible thorough therapy for the damaged brain area, reconstruction of neuronal circuitry, and probable long-term efficacy. Previous studies demonstrated that stem ceils have played crucial roles in various kinds of tissue repair and immune disorders such as cardiovascular diseases, skeletal diseases, arthritis diseases, and lupus erythematodes.37 Evidence from lab experiments and clinical trials implies that adipose-derived stem cells （ADSCs） would be a feasible and ideal cell therapy source because of their multi-differentiation potentials, easy collection, low invasion to donor patients, rapid in vitro amplification, reliable biosafety, non-immunogenicity, privileged immune modulation, and escaping from ethical issues.8-12： Data on neurological disease treatment found that ADSCs transplantation protected against acute inflammatory injury, delayed neuron degeneration, and improved neurological behaviors.12-15 Previously, the detailed mechanisms by which transplanted ADSCs induce functional recovery for stroke treatment are far from being clearly demonstrated. Some proposed reasonable mechanisms just stayed at the level of explanation and hypothesis due to the absence of solid experimental evidence.13-14 In this article, we analyzed the previous data to figure out some new paths or ideas which may do favor to future translational clinical studies.     

Isolation and culture of adult Sertoli cells and their effects on the function of co-cultured allogeneic islets in vitro

Background Globally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered ‘nurse cells’in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro.Methods Adult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets’ function when they were co-cultured for 21 days in vitro.Results In cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P>0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P<0.01).Conclusions A method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets’ function when they were co-cultured in vitro. They could be a helper cell in islet transplantation. Chin Med J 2005; 118(22):1857-1862     

大鼠骨髓间充质干细胞向软骨、脂肪和神经元样细胞分化的研究

目的：探讨体外大鼠骨髓间充质干细胞（rMSCs）多向分化潜能。 方法：从大鼠骨髓中获得间充质干细胞,体外培养传代。通过地塞米松、TGF-β1和维生素C诱导rMSCs向软骨细胞分化,甲苯胺蓝染色和免疫细胞化学检测Ⅱ型胶原进行鉴定。通过地塞米松和胰岛素诱导rMSCs向脂肪细胞分化,采用苏丹Ⅳ染色鉴定。在含IBMX、GDNF和IL-1β的培养基中诱导rMSCs向神经元样细胞分化,免疫细胞化学方法检测神经细胞特异标志NSE和MAP-2a,b。 结果：rMSCs被诱导7 d后,分化成软骨细胞,甲苯胺蓝异染阳性、Ⅱ型胶原阳性。被诱导10 d后,分化成脂肪细胞,苏丹Ⅳ染色阳性。被诱导7 d和15 d后,分化成神经元样细胞,表达NSE和MAP-2a,b,15 d后NSE阳性率是(20.060±0.790)%,MAP-2a,b阳性率是(17.364±5.738)%。 结论：体外rMSCs被诱导分化成软骨细胞、脂肪细胞和神经元样细胞,证明其具有多向分化潜能, 是组织工程研究中最有前途的种子细胞之一。     

当归诱导骨髓间充质干细胞分化为神经元样细胞

目的：研究当归体外定向诱导大鼠骨髓间充质干细胞(MSCs)分化的作用。方法：通过贴壁法分离大鼠MSCs，体外扩增培养。MSC用20μl当归注射液／ml含10％胎牛血清的L—DMEM为预诱导条件，预诱导24h后用当归注射液100μl/ml不含胎牛血清的L—DMEM进行诱导。光镜下观察细胞形态，免疫细胞化学检测神经细胞特异性抗原标志。结果：大鼠骨髓间充质干细胞可通过贴壁法成功分离并可在体外大量扩增。当归诱导3h后大部分MSCs转变为神经元样细胞，出现胞体和突起，免疫细胞化学染色NSE及nestin呈阳性、GFAP性。结论:大鼠骨髓间质干细胞可在体外诱导分化为神经元样细胞。     

脑源性神经营养因子体外诱导骨髓间充质干细胞分化为神经元样细胞

目的探讨脑源性神经营养因子(BDNF)体外诱导骨髓间充质干细胞(MSCs)分化成神经元样细胞的作用及其对诱导后的神经元样细胞保护作用，为治疗神经系统疾病提供更优良的细胞。方法体外培养MSCs至第5代后，分别用BDNF、β-巯基乙醇(β-ME)诱导MSCs，诱导后的第1、3和6小时计算两组神经元样细胞数，对各时间点两组神经元样细胞阳性率进行了比较；诱导后行神经细胞标记抗体的免疫细胞化学、RT-PCR及蛋白质印迹鉴定。结果两组诱导剂诱导后的细胞均呈神经元样细胞的形态；BDNF组诱导分化后细胞的存活时间远较β-ME组长；BDNF组在诱导后6h近于诱导高峰，而β-ME在诱导2h即达高峰，但随后神经元样细胞渐死亡。免疫细胞化学结果显示在两组诱导后细胞的巢蛋白(Nestin)、神经细胞特异性烯醇化酶(NSE)、微管相关蛋白-2(MAP-2)、神经丝蛋白(NF)表达均呈阳性，而胶质纤维酸性蛋白(GFAP)表达呈阴性；RT-PCR显示NSE、NFL，MAP-2的mRNA表达阳性，GFAP有较弱的表达；蛋白质印迹可见两组诱导剂诱导的细胞均表达神经元的特异性标记抗体NSE。结论BDNF能单独诱导MSCs分化形成神经元样细胞．而且分化后细胞的存活时间长．有望用于治疗脑缺血、神经变性等疾病。     

Targeted induction of differentiation of human bone mesenchymal stem cells into neuron-like cells

A systematic method of isolating and culturing human bone mesenchymal stem cells （hMSCs）, and inducing them to differentiate into neuron-like cells in vitro was established. The hMSCs were isolated from bone marrow with the lymphocyte-separating medium, cultured and expanded in vitro, and induced after addition of compound neuro-revulsants. The morphological changes of hMSCs were observed, and the expression of surface markers in induced hMSCs was immunocytochemically identified during induction period. The hMSCs could be separated, cultured and expanded in vitro. After induction by compound neuro-revulsants for 48 h, the changes of neuron-like cells, such as cellular shrinkage and neurite growth, were observed in some cells. The immunochemical staining revealed nestin （＋） or NF （＋）, and GFAP （-）. It was concluded that hMSCs were successfully cultured and induced to differentiate into neuron-like cells.     

鼠胶质瘤细胞上清液诱导人骨髓间充质干细胞向神经元样细胞的分化

目的：分离培养人骨髓间充质干细胞(MSCs),探讨鼠胶质瘤细胞上清液对其向神经元样细胞的诱导分化。方法：利用Percoll梯度分离法,培养人MSCs。采用鼠胶质瘤细胞上清液对MSCs进行诱导分化。观察人MSCs经诱导后细胞的形态变化,采用免疫细胞化学方法检测神经元烯醇化酶(NSE)、神经丝蛋白(NF)、胶质纤维酸性蛋白(GFAP)的表达。 结果：诱导24 h后,细胞胞体收缩呈锥形或球形,有突起长出,细胞间突起相互连接,交错成网,为典型的神经元样形态。免疫细胞化学结果显示,经诱导培养后的神经元样细胞的胞体及部分突起NSE和NF染色呈强阳性表达,而GFAP 染色呈阴性。诱导组出现NSE阳性的细胞率为(79.5±3.2)%,而对照组出现NSE阳性的细胞率为(12.1±2.0)%,两组之间比较差异有显著性 (P<0.01)。诱导组出现NF阳性的细胞率为(41.2±2.4)%,而对照组为阴性。结论：鼠胶质瘤细胞上清液可以诱导MSCs向神经元样细胞分化。     

Adipose tissue-derived stromal cells express neuronal phenotypes

Background Adipose tissue-derived stromal cell (ADSCs) can be greatly expanded in vitro, and induced to differentiate into multiple mesenchymal cell types, including osteogenic, chondrogenic, myogenic, and adipogenic cells. This study was designed to investigate the possibility of ADSCs differentiating into neurons. Methods Adipose tissue from rats was digested with collagenase, and adherent stromal cells were cultured. A medium containing a low concentration of fetal bovine serum was adopted to induce the cells to differentiate. ADSCs were identified by immunocytochemistry, and semi-quantitative RT-PCR was applied to detect mRNA expression of neurofilament 1 (NF1), nestin, and neuron-specific enolase (NSE). Results Nestin-positive cells were found occasionally among ADSCs. ADSCs were found to express NSE mRNA and nestin mRNA, but not NF1 mRNA. ADSCs could differentiate into neuron-like cells in a medium composed of a low concentration of fetal bovine serum, and these differentiated cells displayed complicated neuron-like morphologies. Conclusions The data support the hypothesis that adipose tissue contains stem cells capable of differentiating into neurons. These stem cells can overcome their mesenchymal commitment, and may represent an alternative autologous stem cell source for CNS cell transplantation.     

兔骨髓基质细胞诱导分化成神经元样细胞的研究

OBJECTIVE: To study whether the neuron-like cells derived from bone marrow stromal cells (BMSCs) may excrete amino acids with neurobiological activities and possess the biochemical characteristics of neurons. METHOD: Under sterile condition, BMSCs from New Zealand rabbits were purified by gradient density centrifugation, and were induced to differentiate into neural stem cells and neuronal-like cells in the culture medium for neural stem cells containing retinoic acid (RA, 0.5 microg/ml) and glial-derived neurotrophic factor (GDNF, 20 ng/ml). The differentiated cells were then examined with immunocytochemical method and high-performance liquid chromatograpy (HPLC). RESULTS: The round and enlarged BMSCs on day 10 of cell culture were positive for nestin, and on day 20, the cells with RA+GDNF stimulation differentiated into neuron-like cells with long protrusions and presented neuron-specific enolase (NSE) antigen. HPLC identified high levels of amino acids like Asp, Glu, Gly and Ala in the differentiated cells (P<0.01). CONCLUSIONS: Rabbit BMSCs may proliferate in vitro as from nestin-positive cells and differentiate into NSE-positive cells containing high levels of excitatory and inhibitory amino acid neurotransmitters. RA and GDNF are important promoters for in vitro differentiation of the BMSCs toward neural stem cells.     

大鼠骨髓间充质干细胞体外分化为神经元样细胞的研究

目的：研究大鼠骨髓间充质干细胞(rMSCs)体外扩增不同诱导剂向神经元样细胞诱导分化的潜能。方法：采用贴壁筛选法分离Wistar大鼠股骨rMSCs，体外扩增5-10代后，分别用2-巯基乙醇、硫代甘油等无血清的DMEM培养液诱导hMSCs分化为神经元。用流式细胞仪鉴定rMSCs特异抗原表达，免疫组化鉴定诱导后神经元样细胞的神经元烯醇化酶(NSH)、神经丝蛋白(NF)、胶质纤维酸性蛋白(GFAP)。结果：大鼠MSCs体外扩增传至5代后，流式细胞仪显示rMSCsCD27、CD44、CD90呈阳性表达；将rMSC加入2-巯基乙醇和硫代甘油等诱导剂3h后，rMSC胞体收缩，突起伸出；免疫组化显示诱导出的神经元样细胞NSE、NF表达阳性，GFAP阴性。结论：大鼠骨髓间充质干细胞在体外-定条件下可以分化为神经元样细胞。     

骨髓间充质干细胞向神经元的诱导分化

目的 :探讨大鼠骨髓间充质干细胞 ( rat mesenchymal stem cells,r MSCs)体外诱导分化成神经元的能力。方法 :用 0 .2 5、0 .5、1 .0 mmol· L-1IBMX诱导传代的 r MSCs,待细胞分化后进行形态学观察 ,并用免疫细胞化学方法进行细胞鉴定。结果 :0 .5 mmol· L-1IBMX诱导细胞效果最好 ,2 d即可见有神经元样细胞出现 ,6d神经元样细胞占细胞总数的 ( 2 3.2± 2 .3) %,NSE染色阳性。未分化细胞表达 nestin,诱导 3d,阳性细胞增多 ,6d减少。结论 :体外 r MSC能扩增、传代 ,并被诱导分化成神经元样细胞     

成人骨髓间充质干细胞分化为神经元样细胞的逆转化现象

目的研究成人骨髓间充质干细胞(hMSC)分化为神经元样细胞后的逆转化现象。方法从成人骨髓分离、培养和扩增hMSC。用参芪液诱导hMSC分化为神经元样细胞,并用免疫组化方法鉴定神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF)、胶质纤维酸性蛋白(GFAP)的表达。采用RT-PCR方法检测10个基因(内胚层基因ceruloplasmin;中胚层基因SM22;外胚层基因Amyloidprecursorprotein、syntaxin;生殖系基因protamine;神经元特异性基因NeuroD、NF、NSE、GFAP、Tau)在诱导分化为神经元样细胞的动态变化以及逆转化细胞的基因表达变化。结果hMSC经参芪液诱导后,可见神经元样细胞。免疫组化显示诱导出的神经元样细胞表达NSE、NF阳性,GFAP阴性。去除参芪液,观察到神经元样细胞又恢复为hMSC的外形,呈现宽大扁平或长梭形。基因检测显示逆转化的细胞基因表达与未分化hMSC相似。结论参芪液可以促进hMSC分化为神经元样细胞,诱导分化过程可以出现逆转化现象。