同一个体来源正常与炎性牙周膜干细胞生物学性能的比较

目的:比较同一个体来源正常与炎性牙周膜干细胞的生物学性能。方法:分别分离培养同一个体来源正常牙周膜干细胞(h PDLSCs)及炎性牙周膜干细胞(i-h PDLSCs)。流式细胞仪鉴定两种干细胞表面抗原标记;MTT法、克隆形成率检测并比较两者的增殖能力;茜素红染色及油红O染色检测并比较其分化能力;RT-PCR检测成骨相关基因COL-1、Runx-2、BSP-mRNA及成脂相关基因LPL与PPARγmRNA的表达水平。结果:i-h PDLSCs及h PDLSCs均具有间充质干细胞特性,i-h PDLSCs增殖能力强于h PDLSCs,但分化能力明显弱于h PDLSCs(P<0.05);两者成脂能力比较无统计学差异(P>0.05)。结论:炎症微环境对牙周膜干细胞的生物学性能有一定的影响。

巨噬细胞和淋巴细胞及增殖细胞核抗原在鼻息肉组织中的表达及病理学意义

目的 研究巨噬细胞 (CD68)、T细胞 (CD45RO)、B细胞 (CD2 0 )和增殖核细胞核抗原(proliferatingcellnuclearantigen ,PCNA)在鼻息肉组织中的表达。方法 应用免疫组化链霉菌抗生物素蛋白过氧化酶 (strept avidinoxidase,SP)法对 50例鼻息肉分别做CD2 0、CD45RO、CD68、PCNA的免疫组化染色 ,结合常规HE染色切片进行分析。结果 ①CD68+ 细胞在嗜酸性粒细胞性鼻息肉较嗜中性粒细胞性鼻息肉表达率高 ,差异具有显著性意义 (P <0 0 5) ;②CD45RO、CD2 0在鼻息肉均有阳性表达 ,CD45RO与CD2 0呈负相关 (P =0 0 5) ;③CD68阳性细胞与嗜酸性粒细胞浸润及鼻息肉上皮PCNA阳性表达有相关性 (P <0 0 5)。鼻息肉上皮的PCNA阳性表达和成纤维细胞PCNA阳性表达有相关性 ,(P <0 0 5)。结论 鼻息肉的形成与炎性细胞浸润密切相关。鼻黏膜局部的细胞免疫与体液免疫异常导致上皮细胞、成纤维细胞增殖、腺体增生是鼻息肉发生的基础 ,CD68+

Crohn’s disease-Defect in innate defence

Crohn’s disease may prinicipally involve the whole gastrointestinal tract. Most commonly, the inflammation occurs in the small intestine and/or in the colon with stable disease location over the years. The pathogenesis of both disease phenotypes is complex, the likely primary defect lies in the innate rather than adaptive immunity, particularly in the chemical antimicrobial barrier of the mucosa. Crohn’s ileitis is associated with a reduced expression of the Wnt signalling pathway transcription factor T-cell factor 4 (TCF4), which is regulating Paneth cell differentiation. As a result, the alpha-defensins and principal Paneth cell products HD5 and HD6 are deficiently expressed in ileal disease, independent of current inflammation. In contrast, Crohn’s colitis is typically associated with an impaired induction of the beta-defensins HBD2 and HBD3 caused by fewer gene copy numbers in the gene locus of the beta-defensins on chromosome 8. This ileal and colonic defect in innate defence mediated by a deficiency of the protective alpha- and beta- defensins may enable the luminal microbes to invade the mucosa and trigger the inflammation. A better understanding of the exact molecular mechanisms behind ileal and colonic Crohn’s disease may give rise to new therapeutic strategies based on a stimulation of the protective innate immune system.

Physiological and pathological role of local and immigrating colonic stem cells

The latest avenue of research is revealing the existence of and role for the colonic stem cells in the physiological renewal of the mucosa and in pathological circumstanc- es where they have both positive and negative effects. In the case of human colon, different levels of stem cell compartments exist. First, the crypt epithelial stem cells, which have a role in the normal crypt epithelial cell dynamics and in colorectal carcinogenesis. Close to the crypts, the second layer of stern cells can be found; the local subepithelial stem cell niche, including the pericryptic subepithelial myofibroblasts that regulate the epithelial cell differentiation and have a crucial role in cancer progression and chronic inflammation-related fibrosis. The third level of stem cell compartment is the immigrating bone-marrow-derived stem cells, which have an important role in wound healing after severe mucosal inflammation, but are also involved in cancer invasion. This paper focuses on stem cell biology in the context of physiological and pathological processes in the human colon.

Down-regulation of IL-8 expression in human airway epithelial cells through helper-dependent adenoviral-mediated RNA interference

Interleukin (IL)-8 is a potent neutrophil chemotactic factor and a crucial mediator in neutrophil-dependent inflammation.Various cell types produce IL-8, either in response to external stimuli such as cytokines or bacterial infection, or aftermalignant transformation. Anti-IL-8 strategies have been considered for anti-inflammatory therapy. In this paper wedemonstrate that the RNA interference technique can be used to efficiently down-regulate IL-8 protein expression inairway epithelial cells. We used a helper-dependent adenoviral vector to express a small hairpin (sh)RNA targetinghuman IL-8 in cultured airway epithelial cells (IB3-1, Cftr-/-; C38, Cftr-corrected) stimulated with TNF-α, IL-1β orheat-inactivated Burkholderia cenocepacia. Stimulated IL-8 expression in IB3-1 and C38 cells was significantly reducedby shRNA expression. The shRNA targeting IL-8 had no effect on the activation of NF-κB, or on the protein levels ofIκB or IL-6, suggesting that this anti-IL-8 strategy was highly specific, and therefore may offer potential for thetreatment of inflammatory diseases.

G-CSF in Peg-IFN induced neutropenia in liver transplanted patients with HCV recurrence

AIM:To evaluate the efficacy of granulocyte colony stimulating factors(G-CSF)in liver transplanted patients with hepatitis C(HCV)recurrence and Pegylated-IFN α-2b induced neutropenia,and to evaluate the impact of G-CSF administration on virological response. METHODS:Sixty-eight patients undergoing antiviral treatment for post-liver transplantation(OLT)HCV recurrence were enrolled.All patients developing neutropenia received G-CSF. RESULTS:Twenty three(34%)received G-CSF.Mean neutrophil count at the onset of neutropenia was 700/mmc(range 400-750/mmc);after 1 mo of G-CSF it increased to 1210/mmc(range 300-5590/mmc) (P<0.0001).Three patients did not respond to G-CSF. Treatment duration was similar in neutropenic and non-neutropenic patients.No differences in the rate of discontinuation,infections or virological response were observed between the two groups.G-CSF was protective for the onset of de novo autoimmune hepatitis(P<0.003). CONCLUSION:G-CSF administration is effective in the case of Peg-IFN induced neutropenia increasingneutrophil count,prolonging treatment and leading to sustained virological response(SVR)rates comparable to non-neutropenic patients.It prevents the occurrence of de novo autoimmune hepatitis.

Role of bone marrow-derived mesenchymal stem cells in a rat model of severe acute pancreatitis

AIM:To investigate the role and potential mechanisms of bone marrow mesenchymal stem cells(MSCs) in severe acute peritonitis(SAP).METHODS:Pancreatic acinar cells from Sprague Dawley rats were randomly divided into three groups:nonsodium deoxycholate(SDOC) group(non-SODC group),SDOC group,and a MSCs intervention group(i.e.,a co-culture system of MSCs and pancreatic acinar cells + SDOC).The cell survival rate,the concentration of malonaldehyde(MDA),the density of superoxide dismutase(SOD),serum amylase(AMS) secretion rate and lactate dehydrogenase(LDH) leakage rate were detected at various time points.In a separate study,Sprague Dawley rats were randomly divided into either an SAP group or an SAP + MSCs group.Serum AMS,MDA and SOD,interleukin(IL)-6,IL-10,and tumor necrosis factor(TNF)-α levels,intestinal mucosa injury scores and proliferating cells of small intestinal mucosa were measured at various time points after injecting either MSCs or saline into rats.In both studies,the protective effect of MSCs was evaluated.RESULTS:In vitro,The cell survival rate of pancreatic acinar cells and the density of SOD were significantly reduced,and the concentration of MDA,AMS secretion rate and LDH leakage rate were significantly increased in the SDOC group compared with the MSCs intervention group and the Non-SDOC group at each time point.In vivo,Serum AMS,IL-6,TNF-α and MAD level in the SAP + MSCs group were lower than the SAP group;however serum IL-10 level was higher than the SAP group.Serum SOD level was higher than the SAP group at each time point,whereas a significant betweengroup difference in SOD level was only noted after 24 h.Intestinal mucosa injury scores was significantly reduced and the proliferating cells of small intestinal mucosa became obvious after injecting MSCs.CONCLUSION:MSCs can effectively relieve injury to pancreatic acinar cells and small intestinal epithelium,promote the proliferation of enteric epithelium and repair of the mucosa,attenuate systemic inflammation in rats with SAP.

Regulatory effect of moxibustion for rats with ulcerative colitis on the macrophage functional phenotype protein of lung tissueRegulatory effect of moxibustion for rats with ulcerative colitis on the macrophage functional phenotype protein of lung tissue

Objective： To observe the regulatory effect of moxibustion on the expression of CD86 and CD163, which are the important functional phenotypes of macrophage differentiation in lung tissue of ulcerative colitis（UC） rats. The mechanism of the regulatory effect of moxibustion for macrophage differentiation based on the key cytokine interferon-gamma（IFN-γ）, tumor necrosis death factor-alpha（TNF-α）, interleukin- 4（IL-4） and interleukin-13（IL-13） was also explored. Methods： Forty SD rats were randomly divided into a normal group（NG）, a model group（MG）, a normal moxibustion group（NMG） and a smokeless moxibustion group（SMG）. The model of UC was made by antigen immunization combined with enema with topical formalin. The rats in the normal moxibustion group accepted moxibustion at bilateral Tianshu（ST 25）, while the rats in the smokeless moxibustion group with smokeless moxibustion at bilateral Tianshu（ST 25）, each 10 min, once a day for 8 d. After treatments, the hematoxylin-eosin（HE） staining was used to observe the pathological changes of colonic tissue, the Western blotting（WB） was used to observe the expression of CD86 and CD163, two important functional phenotypes of macrophage differentiation in lung tissue of UC rats, while the enzyme-linked immunosorbent assay（ELISA） was used to assess the content of IFN-γ, TNF-α, IL-4 and IL-13, the key cytokines of macrophage differentiation in micro environment of the lung tissue. Results： Compared with the NG, the colons of rats in MG were injured more seriously, and the scores of gross observation and histological examination were significantly higher（P〈0.05）. Compared with the MG, the pathological changes of the two groups of rats with moxibustion treatment were improved, which presented with ulcer repair, inflammation dissipation, and the general score and histological score of the two groups were decreased（P〈0.05）. Compared with the NG, the expression of CD86 in the lung tissue of rats in the MG was increased（P〈0.05）, and CD163 expression was decreased（P〈0.05）. Compared with the MG and the SMG, the expression of CD86 in the lung tissue of the rats in the NMG was significantly lower than those in the MG and SMG, and CD163 was higher（P〈0.05）, while the differences were not statistically significant between the MG and the SMG（P〈0.05）. Compared with the NG, the expression of key cytokines in lung tissue of MG was abnormal, the contents of IFN-γ and TNF-α increased（P〈0.05）, while the IL-4 and IL-13 decreased（P〈0.05）. The IL-4 and IL-13 were significantly increased in lung tissue of rats in the NMG（P〈0.05）, and the IFN-γ and TNF-α were reduced（P〈0.05）, compared with those in the MG and the SMG, while the differences were not statistically significant between the MG and the SMG（P〈0.05）. Conclusion： Moxibustion can increase the expression of CD163, the important functional phenotype of macrophages in lung tissue of UC rats, and the differentiation critical cytokines IL-4 and IL-13. It can also reduce activated phenotype CD86 and its differentiation critical cytokines IFN-γ and TNF-α in the lung tissue of UC rats.

Effect of Ermiao Fang with Xixin(Herba Asari Mandshurici) on bone marrow stem cell directional homing to a focal zone in an osteoarthritis rat model

OBJECTIVE: To investigate the effects of Ermiao Fang(EM) with medical guide Xixin(Herba Asari Mandshurici)(HAM) on bone marrow stem cell migration to a focal zone in osteoarthritis(OA) rats.METHODS: OA rats were induced by arthrectomy and assigned to sham-operated, model, EM, or EM plus HAM groups.All rats were injected with recombinant human granulocyte colony-stimulating factor 30μg·kg-1·d-1for7 days and treated with EMor EM plus HAM at 1.6 or 1.9 g·kg-1·d-1 for 3 or 6 weeks, respectively. Chondrocyte apoptosis and cartilage matrix components were tested by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay and special staining. Levels of interleukin-1 beta(IL-1β) tumor necrosis factor alpha(TNF-α) nitric oxide(NO), and inducible nitric oxide synthase(iNOS) in serum were detected by enzyme-linked immunosorbent assay or radioimmunoassay. Matrix metalloproteinases(MMPs)-13,tissue inhibitors of metalloproteinases(TIMPs)-1,Bromodeoxyuridine(BrdU), cluster of differentiation 34(CD34), and stromal cell-derived factor 1(SDF-1) were measured by immunohistochemical assay.RESULTS:The EM and EM plus HAM groups had significantly less cartilage damage and synovium inflammation the model group. Moreover, the EM and EM plus HAM groups had less chondrocyte apoptosis and more proteoglycan and collagen content than the model group.The EM and EMplus HAM groups had obviously higher MMPs-13 and TIMPs-1 expression in the cartilage than the model group. Moreover, the two formula groups had less release of IL-1β, TNF-α, NO, and iNOS than model group. Importantly, the expressions of BrdU, CD34,and SDF-1 in cartilage were significantly higher in the EM and EM plus HAM-Medtreated rats than model group. Notably, the EM plus HAM treatment seemed to have the greatest effects.CONCLUSION: HAM improves the therapeutic effects of EM on OA rats by enhancing BMSC directional homing to the focal zone.